ERS4855907 SAMEA7095329 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095329 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#35 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#35 scientific_name Mus musculus strain C57BL/6J ERS4855908 SAMEA7095330 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095330 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#350 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#350 scientific_name Mus musculus strain C57BL/6J ERS4855909 SAMEA7095331 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095331 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#351 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#351 scientific_name Mus musculus strain C57BL/6J ERS4855910 SAMEA7095332 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095332 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#352 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#352 scientific_name Mus musculus strain C57BL/6J ERS4855911 SAMEA7095333 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095333 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#353 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#353 scientific_name Mus musculus strain C57BL/6J ERS4855912 SAMEA7095334 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095334 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#354 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#354 scientific_name Mus musculus strain C57BL/6J ERS4855913 SAMEA7095335 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095335 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#355 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#355 scientific_name Mus musculus strain C57BL/6J ERS4855914 SAMEA7095336 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095336 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#356 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#356 scientific_name Mus musculus strain C57BL/6J ERS4855915 SAMEA7095337 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095337 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#357 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#357 scientific_name Mus musculus strain C57BL/6J ERS4855916 SAMEA7095338 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095338 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#358 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#358 scientific_name Mus musculus strain C57BL/6J ERS4855917 SAMEA7095339 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095339 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#359 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#359 scientific_name Mus musculus strain C57BL/6J ERS4855918 SAMEA7095340 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095340 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#36 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#36 scientific_name Mus musculus strain C57BL/6J ERS4855919 SAMEA7095341 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095341 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#360 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#360 scientific_name Mus musculus strain C57BL/6J ERS4855920 SAMEA7095342 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095342 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#361 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:27921_6#361 scientific_name Mus musculus strain C57BL/6J ERS4855921 SAMEA7095343 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095343 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#362 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:27921_6#362 scientific_name Mus musculus strain C57BL/6J ERS4855922 SAMEA7095344 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095344 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#363 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:27921_6#363 scientific_name Mus musculus strain C57BL/6J ERS4855923 SAMEA7095345 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095345 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#364 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:27921_6#364 scientific_name Mus musculus strain C57BL/6J ERS4855924 SAMEA7095346 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095346 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#365 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:27921_6#365 scientific_name Mus musculus strain C57BL/6J ERS4855925 SAMEA7095347 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095347 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#366 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:27921_6#366 scientific_name Mus musculus strain C57BL/6J ERS4855926 SAMEA7095348 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095348 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#367 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:27921_6#367 scientific_name Mus musculus strain C57BL/6J ERS4855927 SAMEA7095349 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095349 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#368 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:27921_6#368 scientific_name Mus musculus strain C57BL/6J ERS4855928 SAMEA7095350 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095350 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#369 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:27921_6#369 scientific_name Mus musculus strain C57BL/6J ERS4855929 SAMEA7095351 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095351 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#37 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#37 scientific_name Mus musculus strain C57BL/6J ERS4855930 SAMEA7095352 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095352 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#370 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:27921_6#370 scientific_name Mus musculus strain C57BL/6J ERS4855931 SAMEA7095353 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095353 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#371 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:27921_6#371 scientific_name Mus musculus strain C57BL/6J ERS4855932 SAMEA7095354 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095354 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#372 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:27921_6#372 scientific_name Mus musculus strain C57BL/6J ERS4855933 SAMEA7095355 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095355 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#373 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#373 scientific_name Mus musculus strain C57BL/6J ERS4855934 SAMEA7095356 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095356 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#374 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#374 scientific_name Mus musculus strain C57BL/6J ERS4855935 SAMEA7095357 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095357 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#375 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#375 scientific_name Mus musculus strain C57BL/6J ERS4855936 SAMEA7095358 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095358 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#376 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#376 scientific_name Mus musculus strain C57BL/6J ERS4855937 SAMEA7095359 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095359 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#377 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#377 scientific_name Mus musculus strain C57BL/6J ERS4855938 SAMEA7095360 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095360 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#378 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#378 scientific_name Mus musculus strain C57BL/6J ERS4855939 SAMEA7095361 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095361 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#379 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#379 scientific_name Mus musculus strain C57BL/6J ERS4855940 SAMEA7095362 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095362 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#38 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#38 scientific_name Mus musculus strain C57BL/6J ERS4855941 SAMEA7095363 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:44Z External Id SAMEA7095363 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:44Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#380 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#380 scientific_name Mus musculus strain C57BL/6J ERS4855942 SAMEA7095364 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095364 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#381 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#381 scientific_name Mus musculus strain C57BL/6J ERS4855943 SAMEA7095365 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095365 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#382 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#382 scientific_name Mus musculus strain C57BL/6J ERS4855944 SAMEA7095366 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095366 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#383 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#383 scientific_name Mus musculus strain C57BL/6J ERS4855945 SAMEA7095367 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095367 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#384 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:27921_6#384 scientific_name Mus musculus strain C57BL/6J ERS4855946 SAMEA7095368 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095368 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#39 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#39 scientific_name Mus musculus strain C57BL/6J ERS4855947 SAMEA7095369 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095369 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#4 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#4 scientific_name Mus musculus strain C57BL/6J ERS4855948 SAMEA7095370 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095370 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#40 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#40 scientific_name Mus musculus strain C57BL/6J ERS4855949 SAMEA7095371 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095371 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#41 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#41 scientific_name Mus musculus strain C57BL/6J ERS4855950 SAMEA7095372 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095372 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#42 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#42 scientific_name Mus musculus strain C57BL/6J ERS4855951 SAMEA7095373 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095373 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#43 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#43 scientific_name Mus musculus strain C57BL/6J ERS4855952 SAMEA7095374 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095374 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#44 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#44 scientific_name Mus musculus strain C57BL/6J ERS4855953 SAMEA7095375 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095375 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#45 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#45 scientific_name Mus musculus strain C57BL/6J ERS4855954 SAMEA7095376 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095376 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#46 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#46 scientific_name Mus musculus strain C57BL/6J ERS4855955 SAMEA7095377 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095377 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#47 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#47 scientific_name Mus musculus strain C57BL/6J ERS4855956 SAMEA7095378 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095378 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#48 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#48 scientific_name Mus musculus strain C57BL/6J ERS4855957 SAMEA7095379 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095379 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#49 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#49 scientific_name Mus musculus strain C57BL/6J ERS4855958 SAMEA7095380 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095380 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#5 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#5 scientific_name Mus musculus strain C57BL/6J ERS4855959 SAMEA7095381 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095381 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#50 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#50 scientific_name Mus musculus strain C57BL/6J ERS4855960 SAMEA7095382 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095382 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#51 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#51 scientific_name Mus musculus strain C57BL/6J ERS4855961 SAMEA7095383 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095383 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#52 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#52 scientific_name Mus musculus strain C57BL/6J ERS4855962 SAMEA7095384 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095384 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#53 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#53 scientific_name Mus musculus strain C57BL/6J ERS4855963 SAMEA7095385 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095385 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#54 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#54 scientific_name Mus musculus strain C57BL/6J ERS4855964 SAMEA7095386 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095386 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#55 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#55 scientific_name Mus musculus strain C57BL/6J ERS4855965 SAMEA7095387 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095387 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#56 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#56 scientific_name Mus musculus strain C57BL/6J ERS4855966 SAMEA7095388 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095388 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#57 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#57 scientific_name Mus musculus strain C57BL/6J ERS4855967 SAMEA7095389 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095389 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#58 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#58 scientific_name Mus musculus strain C57BL/6J ERS4855968 SAMEA7095390 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095390 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#59 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#59 scientific_name Mus musculus strain C57BL/6J ERS4855969 SAMEA7095391 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095391 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#6 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#6 scientific_name Mus musculus strain C57BL/6J ERS4855970 SAMEA7095392 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095392 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#60 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#60 scientific_name Mus musculus strain C57BL/6J ERS4855971 SAMEA7095393 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095393 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#61 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#61 scientific_name Mus musculus strain C57BL/6J ERS4855972 SAMEA7095394 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095394 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#62 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#62 scientific_name Mus musculus strain C57BL/6J ERS4855973 SAMEA7095395 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095395 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#63 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#63 scientific_name Mus musculus strain C57BL/6J ERS4855974 SAMEA7095396 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095396 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#64 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#64 scientific_name Mus musculus strain C57BL/6J ERS4855975 SAMEA7095397 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095397 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#65 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#65 scientific_name Mus musculus strain C57BL/6J ERS4855976 SAMEA7095398 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095398 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#66 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#66 scientific_name Mus musculus strain C57BL/6J ERS4855977 SAMEA7095399 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095399 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#67 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#67 scientific_name Mus musculus strain C57BL/6J ERS4855978 SAMEA7095400 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095400 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#68 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#68 scientific_name Mus musculus strain C57BL/6J ERS4855979 SAMEA7095401 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095401 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#69 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#69 scientific_name Mus musculus strain C57BL/6J ERS4855980 SAMEA7095402 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095402 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#7 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#7 scientific_name Mus musculus strain C57BL/6J ERS4855981 SAMEA7095403 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095403 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#70 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#70 scientific_name Mus musculus strain C57BL/6J ERS4855982 SAMEA7095404 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095404 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#71 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#71 scientific_name Mus musculus strain C57BL/6J ERS4855983 SAMEA7095405 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095405 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#72 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#72 scientific_name Mus musculus strain C57BL/6J ERS4855984 SAMEA7095406 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095406 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#73 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#73 scientific_name Mus musculus strain C57BL/6J ERS4855985 SAMEA7095407 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095407 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#74 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#74 scientific_name Mus musculus strain C57BL/6J ERS4855986 SAMEA7095408 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095408 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#75 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#75 scientific_name Mus musculus strain C57BL/6J ERS4855987 SAMEA7095409 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095409 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#76 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#76 scientific_name Mus musculus strain C57BL/6J ERS4855988 SAMEA7095410 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095410 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#77 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#77 scientific_name Mus musculus strain C57BL/6J ERS4855989 SAMEA7095411 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095411 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#78 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#78 scientific_name Mus musculus strain C57BL/6J ERS4855990 SAMEA7095412 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095412 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#79 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#79 scientific_name Mus musculus strain C57BL/6J ERS4855991 SAMEA7095413 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095413 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#8 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#8 scientific_name Mus musculus strain C57BL/6J ERS4855992 SAMEA7095414 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095414 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#80 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#80 scientific_name Mus musculus strain C57BL/6J ERS4855993 SAMEA7095415 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095415 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#81 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#81 scientific_name Mus musculus strain C57BL/6J ERS4855994 SAMEA7095416 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095416 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#82 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#82 scientific_name Mus musculus strain C57BL/6J ERS4855995 SAMEA7095417 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095417 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#83 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#83 scientific_name Mus musculus strain C57BL/6J ERS4855996 SAMEA7095419 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095419 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#84 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#84 scientific_name Mus musculus strain C57BL/6J ERS4855997 SAMEA7095420 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095420 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#85 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#85 scientific_name Mus musculus strain C57BL/6J ERS4855998 SAMEA7095421 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095421 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#86 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#86 scientific_name Mus musculus strain C57BL/6J ERS4855999 SAMEA7095422 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095422 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#87 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#87 scientific_name Mus musculus strain C57BL/6J ERS4856000 SAMEA7095423 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095423 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#88 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#88 scientific_name Mus musculus strain C57BL/6J ERS4856001 SAMEA7095424 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095424 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#89 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#89 scientific_name Mus musculus strain C57BL/6J ERS4856002 SAMEA7095425 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095425 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#9 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#9 scientific_name Mus musculus strain C57BL/6J ERS4856003 SAMEA7095426 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095426 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#90 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#90 scientific_name Mus musculus strain C57BL/6J ERS4856004 SAMEA7095427 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095427 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#91 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#91 scientific_name Mus musculus strain C57BL/6J ERS4856005 SAMEA7095428 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095428 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#92 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#92 scientific_name Mus musculus strain C57BL/6J ERS4856006 SAMEA7095429 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095429 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#93 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#93 scientific_name Mus musculus strain C57BL/6J ERS4856007 SAMEA7095430 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095430 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#94 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#94 scientific_name Mus musculus strain C57BL/6J ERS4856008 SAMEA7095431 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095431 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#95 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#95 scientific_name Mus musculus strain C57BL/6J ERS4856009 SAMEA7095432 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095432 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#96 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:27921_6#96 scientific_name Mus musculus strain C57BL/6J ERS4856010 SAMEA7095433 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095433 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#97 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#97 scientific_name Mus musculus strain C57BL/6J ERS4856011 SAMEA7095434 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095434 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#98 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#98 scientific_name Mus musculus strain C57BL/6J ERS4856012 SAMEA7095435 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095435 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:27921_6#99 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:27921_6#99 scientific_name Mus musculus strain C57BL/6J ERS4856013 SAMEA7095436 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095436 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#1 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#1 scientific_name Mus musculus strain C57BL/6J ERS4856014 SAMEA7095437 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095437 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#10 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#10 scientific_name Mus musculus strain C57BL/6J ERS4856015 SAMEA7095438 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095438 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#100 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#100 scientific_name Mus musculus strain C57BL/6J ERS4856016 SAMEA7095439 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095439 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#101 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#101 scientific_name Mus musculus strain C57BL/6J ERS4856017 SAMEA7095440 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095440 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#102 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#102 scientific_name Mus musculus strain C57BL/6J ERS4856018 SAMEA7095441 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095441 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#103 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#103 scientific_name Mus musculus strain C57BL/6J ERS4856019 SAMEA7095442 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095442 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#104 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#104 scientific_name Mus musculus strain C57BL/6J ERS4856020 SAMEA7095443 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095443 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#105 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#105 scientific_name Mus musculus strain C57BL/6J ERS4856021 SAMEA7095444 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095444 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#106 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#106 scientific_name Mus musculus strain C57BL/6J ERS4856022 SAMEA7095445 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:45Z External Id SAMEA7095445 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:45Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#107 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#107 scientific_name Mus musculus strain C57BL/6J ERS4856023 SAMEA7095446 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095446 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#108 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#108 scientific_name Mus musculus strain C57BL/6J ERS4856024 SAMEA7095447 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095447 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#109 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#109 scientific_name Mus musculus strain C57BL/6J ERS4856025 SAMEA7095448 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095448 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#11 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#11 scientific_name Mus musculus strain C57BL/6J ERS4856026 SAMEA7095449 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095449 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#110 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#110 scientific_name Mus musculus strain C57BL/6J ERS4856027 SAMEA7095450 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095450 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#111 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#111 scientific_name Mus musculus strain C57BL/6J ERS4856028 SAMEA7095451 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095451 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#112 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#112 scientific_name Mus musculus strain C57BL/6J ERS4856029 SAMEA7095452 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095452 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#113 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#113 scientific_name Mus musculus strain C57BL/6J ERS4856030 SAMEA7095453 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095453 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#114 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#114 scientific_name Mus musculus strain C57BL/6J ERS4856031 SAMEA7095454 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095454 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#115 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#115 scientific_name Mus musculus strain C57BL/6J ERS4856032 SAMEA7095455 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095455 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#116 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#116 scientific_name Mus musculus strain C57BL/6J ERS4856033 SAMEA7095456 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095456 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#117 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#117 scientific_name Mus musculus strain C57BL/6J ERS4856034 SAMEA7095457 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095457 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#118 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#118 scientific_name Mus musculus strain C57BL/6J ERS4856035 SAMEA7095458 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095458 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#119 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#119 scientific_name Mus musculus strain C57BL/6J ERS4856036 SAMEA7095459 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095459 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#12 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#12 scientific_name Mus musculus strain C57BL/6J ERS4856037 SAMEA7095460 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095460 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#120 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#120 scientific_name Mus musculus strain C57BL/6J ERS4856038 SAMEA7095461 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095461 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#121 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#121 scientific_name Mus musculus strain C57BL/6J ERS4856039 SAMEA7095462 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095462 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#122 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#122 scientific_name Mus musculus strain C57BL/6J ERS4856040 SAMEA7095463 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095463 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#123 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#123 scientific_name Mus musculus strain C57BL/6J ERS4856041 SAMEA7095464 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095464 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#124 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#124 scientific_name Mus musculus strain C57BL/6J ERS4856042 SAMEA7095465 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095465 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#125 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#125 scientific_name Mus musculus strain C57BL/6J ERS4856043 SAMEA7095466 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095466 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#126 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#126 scientific_name Mus musculus strain C57BL/6J ERS4856044 SAMEA7095467 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095467 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#127 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#127 scientific_name Mus musculus strain C57BL/6J ERS4856045 SAMEA7095468 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095468 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#128 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#128 scientific_name Mus musculus strain C57BL/6J ERS4856046 SAMEA7095469 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095469 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#129 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#129 scientific_name Mus musculus strain C57BL/6J ERS4856047 SAMEA7095470 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095470 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#13 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#13 scientific_name Mus musculus strain C57BL/6J ERS4856048 SAMEA7095471 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095471 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#130 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#130 scientific_name Mus musculus strain C57BL/6J ERS4856049 SAMEA7095472 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095472 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#131 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#131 scientific_name Mus musculus strain C57BL/6J ERS4856050 SAMEA7095473 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095473 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#132 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#132 scientific_name Mus musculus strain C57BL/6J ERS4856051 SAMEA7095474 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095474 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#133 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#133 scientific_name Mus musculus strain C57BL/6J ERS4856052 SAMEA7095475 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095475 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#134 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#134 scientific_name Mus musculus strain C57BL/6J ERS4856053 SAMEA7095476 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095476 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#135 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#135 scientific_name Mus musculus strain C57BL/6J ERS4856054 SAMEA7095477 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095477 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#136 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#136 scientific_name Mus musculus strain C57BL/6J ERS4856055 SAMEA7095478 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095478 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#137 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#137 scientific_name Mus musculus strain C57BL/6J ERS4856056 SAMEA7095479 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095479 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#138 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#138 scientific_name Mus musculus strain C57BL/6J ERS4856057 SAMEA7095480 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095480 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#139 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#139 scientific_name Mus musculus strain C57BL/6J ERS4856058 SAMEA7095481 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095481 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#14 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#14 scientific_name Mus musculus strain C57BL/6J ERS4856059 SAMEA7095482 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095482 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#140 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#140 scientific_name Mus musculus strain C57BL/6J ERS4856060 SAMEA7095483 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095483 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#141 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#141 scientific_name Mus musculus strain C57BL/6J ERS4856061 SAMEA7095484 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095484 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#142 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#142 scientific_name Mus musculus strain C57BL/6J ERS4856062 SAMEA7095485 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095485 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#143 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#143 scientific_name Mus musculus strain C57BL/6J ERS4856063 SAMEA7095486 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095486 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#144 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#144 scientific_name Mus musculus strain C57BL/6J ERS4856064 SAMEA7095487 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095487 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#145 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#145 scientific_name Mus musculus strain C57BL/6J ERS4856065 SAMEA7095488 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095488 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#146 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#146 scientific_name Mus musculus strain C57BL/6J ERS4856066 SAMEA7095489 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095489 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#147 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#147 scientific_name Mus musculus strain C57BL/6J ERS4856067 SAMEA7095490 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095490 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#148 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#148 scientific_name Mus musculus strain C57BL/6J ERS4856068 SAMEA7095491 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095491 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#149 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#149 scientific_name Mus musculus strain C57BL/6J ERS4856069 SAMEA7095492 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095492 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#15 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#15 scientific_name Mus musculus strain C57BL/6J ERS4856070 SAMEA7095493 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095493 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#150 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#150 scientific_name Mus musculus strain C57BL/6J ERS4856071 SAMEA7095494 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095494 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#151 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#151 scientific_name Mus musculus strain C57BL/6J ERS4856072 SAMEA7095495 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095495 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#152 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#152 scientific_name Mus musculus strain C57BL/6J ERS4856073 SAMEA7095496 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095496 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#153 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#153 scientific_name Mus musculus strain C57BL/6J ERS4856074 SAMEA7095497 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095497 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#154 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#154 scientific_name Mus musculus strain C57BL/6J ERS4856075 SAMEA7095498 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095498 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#155 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#155 scientific_name Mus musculus strain C57BL/6J ERS4856076 SAMEA7095499 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095499 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#156 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#156 scientific_name Mus musculus strain C57BL/6J ERS4856077 SAMEA7095500 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095500 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#157 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#157 scientific_name Mus musculus strain C57BL/6J ERS4856078 SAMEA7095501 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095501 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#158 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#158 scientific_name Mus musculus strain C57BL/6J ERS4856079 SAMEA7095502 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095502 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#159 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#159 scientific_name Mus musculus strain C57BL/6J ERS4856080 SAMEA7095503 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095503 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#16 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#16 scientific_name Mus musculus strain C57BL/6J ERS4856081 SAMEA7095504 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095504 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#160 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#160 scientific_name Mus musculus strain C57BL/6J ERS4856082 SAMEA7095505 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095505 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#161 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#161 scientific_name Mus musculus strain C57BL/6J ERS4856083 SAMEA7095506 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095506 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#162 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#162 scientific_name Mus musculus strain C57BL/6J ERS4856084 SAMEA7095507 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095507 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#163 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#163 scientific_name Mus musculus strain C57BL/6J ERS4856085 SAMEA7095508 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095508 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#164 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#164 scientific_name Mus musculus strain C57BL/6J ERS4856086 SAMEA7095509 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095509 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#165 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#165 scientific_name Mus musculus strain C57BL/6J ERS4856087 SAMEA7095510 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095510 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#166 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#166 scientific_name Mus musculus strain C57BL/6J ERS4856088 SAMEA7095511 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095511 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#167 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#167 scientific_name Mus musculus strain C57BL/6J ERS4856089 SAMEA7095512 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095512 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#169 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#169 scientific_name Mus musculus strain C57BL/6J ERS4856090 SAMEA7095513 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095513 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#17 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#17 scientific_name Mus musculus strain C57BL/6J ERS4856091 SAMEA7095514 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095514 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#170 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#170 scientific_name Mus musculus strain C57BL/6J ERS4856092 SAMEA7095515 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095515 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#171 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#171 scientific_name Mus musculus strain C57BL/6J ERS4856093 SAMEA7095516 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095516 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#172 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#172 scientific_name Mus musculus strain C57BL/6J ERS4856094 SAMEA7095517 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095517 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#173 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#173 scientific_name Mus musculus strain C57BL/6J ERS4856095 SAMEA7095518 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095518 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#174 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#174 scientific_name Mus musculus strain C57BL/6J ERS4856096 SAMEA7095519 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095519 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#175 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#175 scientific_name Mus musculus strain C57BL/6J ERS4856097 SAMEA7095520 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095520 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#176 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#176 scientific_name Mus musculus strain C57BL/6J ERS4856098 SAMEA7095521 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095521 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#177 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#177 scientific_name Mus musculus strain C57BL/6J ERS4856099 SAMEA7095522 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095522 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#178 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#178 scientific_name Mus musculus strain C57BL/6J ERS4856100 SAMEA7095523 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095523 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#179 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#179 scientific_name Mus musculus strain C57BL/6J ERS4856101 SAMEA7095524 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095524 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#18 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#18 scientific_name Mus musculus strain C57BL/6J ERS4856102 SAMEA7095525 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095525 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#180 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#180 scientific_name Mus musculus strain C57BL/6J ERS4856103 SAMEA7095526 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:46Z External Id SAMEA7095526 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:46Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#181 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#181 scientific_name Mus musculus strain C57BL/6J ERS4856104 SAMEA7095527 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095527 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#182 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#182 scientific_name Mus musculus strain C57BL/6J ERS4856105 SAMEA7095528 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095528 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#183 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#183 scientific_name Mus musculus strain C57BL/6J ERS4856106 SAMEA7095529 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095529 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#184 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#184 scientific_name Mus musculus strain C57BL/6J ERS4856107 SAMEA7095530 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095530 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#185 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#185 scientific_name Mus musculus strain C57BL/6J ERS4856108 SAMEA7095531 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095531 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#186 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#186 scientific_name Mus musculus strain C57BL/6J ERS4856109 SAMEA7095532 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095532 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#187 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#187 scientific_name Mus musculus strain C57BL/6J ERS4856110 SAMEA7095533 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095533 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#188 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#188 scientific_name Mus musculus strain C57BL/6J ERS4856111 SAMEA7095534 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095534 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#189 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#189 scientific_name Mus musculus strain C57BL/6J ERS4856112 SAMEA7095535 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095535 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#19 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#19 scientific_name Mus musculus strain C57BL/6J ERS4856113 SAMEA7095536 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095536 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#190 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#190 scientific_name Mus musculus strain C57BL/6J ERS4856114 SAMEA7095537 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095537 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#191 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#191 scientific_name Mus musculus strain C57BL/6J ERS4856115 SAMEA7095538 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095538 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#192 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#192 scientific_name Mus musculus strain C57BL/6J ERS4856116 SAMEA7095539 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095539 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#193 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#193 scientific_name Mus musculus strain C57BL/6J ERS4856117 SAMEA7095540 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095540 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#194 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#194 scientific_name Mus musculus strain C57BL/6J ERS4856118 SAMEA7095541 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095541 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#195 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#195 scientific_name Mus musculus strain C57BL/6J ERS4856119 SAMEA7095542 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095542 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#196 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#196 scientific_name Mus musculus strain C57BL/6J ERS4856120 SAMEA7095543 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095543 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#197 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#197 scientific_name Mus musculus strain C57BL/6J ERS4856121 SAMEA7095544 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095544 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#198 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#198 scientific_name Mus musculus strain C57BL/6J ERS4856122 SAMEA7095545 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095545 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#199 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#199 scientific_name Mus musculus strain C57BL/6J ERS4856123 SAMEA7095546 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095546 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#2 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#2 scientific_name Mus musculus strain C57BL/6J ERS4856124 SAMEA7095547 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095547 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#20 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#20 scientific_name Mus musculus strain C57BL/6J ERS4856125 SAMEA7095548 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095548 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#200 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#200 scientific_name Mus musculus strain C57BL/6J ERS4856126 SAMEA7095549 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095549 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#201 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#201 scientific_name Mus musculus strain C57BL/6J ERS4856127 SAMEA7095550 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095550 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#202 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#202 scientific_name Mus musculus strain C57BL/6J ERS4856128 SAMEA7095551 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095551 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#203 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#203 scientific_name Mus musculus strain C57BL/6J ERS4856129 SAMEA7095552 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095552 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#204 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#204 scientific_name Mus musculus strain C57BL/6J ERS4856130 SAMEA7095553 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095553 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#205 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#205 scientific_name Mus musculus strain C57BL/6J ERS4856131 SAMEA7095554 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095554 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#206 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#206 scientific_name Mus musculus strain C57BL/6J ERS4856132 SAMEA7095555 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095555 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#207 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#207 scientific_name Mus musculus strain C57BL/6J ERS4856133 SAMEA7095556 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095556 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#208 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#208 scientific_name Mus musculus strain C57BL/6J ERS4856134 SAMEA7095557 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095557 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#209 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#209 scientific_name Mus musculus strain C57BL/6J ERS4856135 SAMEA7095558 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095558 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#21 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#21 scientific_name Mus musculus strain C57BL/6J ERS4856136 SAMEA7095559 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095559 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#210 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#210 scientific_name Mus musculus strain C57BL/6J ERS4856137 SAMEA7095560 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095560 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#211 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#211 scientific_name Mus musculus strain C57BL/6J ERS4856138 SAMEA7095561 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095561 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#212 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#212 scientific_name Mus musculus strain C57BL/6J ERS4856139 SAMEA7095562 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095562 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#213 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#213 scientific_name Mus musculus strain C57BL/6J ERS4856140 SAMEA7095563 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095563 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#214 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#214 scientific_name Mus musculus strain C57BL/6J ERS4856141 SAMEA7095564 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095564 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#215 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#215 scientific_name Mus musculus strain C57BL/6J ERS4856142 SAMEA7095565 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095565 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#216 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#216 scientific_name Mus musculus strain C57BL/6J ERS4856143 SAMEA7095566 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095566 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#217 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#217 scientific_name Mus musculus strain C57BL/6J ERS4856144 SAMEA7095567 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095567 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#218 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#218 scientific_name Mus musculus strain C57BL/6J ERS4856145 SAMEA7095568 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095568 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#219 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#219 scientific_name Mus musculus strain C57BL/6J ERS4856146 SAMEA7095569 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095569 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#22 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#22 scientific_name Mus musculus strain C57BL/6J ERS4856147 SAMEA7095570 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095570 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#220 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#220 scientific_name Mus musculus strain C57BL/6J ERS4856148 SAMEA7095571 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095571 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#221 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#221 scientific_name Mus musculus strain C57BL/6J ERS4856149 SAMEA7095572 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095572 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#222 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#222 scientific_name Mus musculus strain C57BL/6J ERS4856150 SAMEA7095573 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095573 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#223 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#223 scientific_name Mus musculus strain C57BL/6J ERS4856151 SAMEA7095574 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095574 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#224 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#224 scientific_name Mus musculus strain C57BL/6J ERS4856152 SAMEA7095575 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095575 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#225 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#225 scientific_name Mus musculus strain C57BL/6J ERS4856153 SAMEA7095576 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095576 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#226 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#226 scientific_name Mus musculus strain C57BL/6J ERS4856154 SAMEA7095577 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095577 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#227 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#227 scientific_name Mus musculus strain C57BL/6J ERS4856155 SAMEA7095578 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095578 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#228 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#228 scientific_name Mus musculus strain C57BL/6J ERS4856156 SAMEA7095579 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095579 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#229 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#229 scientific_name Mus musculus strain C57BL/6J ERS4856157 SAMEA7095580 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095580 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#23 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#23 scientific_name Mus musculus strain C57BL/6J ERS4856158 SAMEA7095581 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095581 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#230 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#230 scientific_name Mus musculus strain C57BL/6J ERS4856159 SAMEA7095582 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095582 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#231 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#231 scientific_name Mus musculus strain C57BL/6J ERS4856160 SAMEA7095583 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095583 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#232 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#232 scientific_name Mus musculus strain C57BL/6J ERS4856161 SAMEA7095584 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095584 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#233 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#233 scientific_name Mus musculus strain C57BL/6J ERS4856162 SAMEA7095585 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095585 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#234 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#234 scientific_name Mus musculus strain C57BL/6J ERS4856163 SAMEA7095586 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095586 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#235 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#235 scientific_name Mus musculus strain C57BL/6J ERS4856164 SAMEA7095587 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095587 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#236 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#236 scientific_name Mus musculus strain C57BL/6J ERS4856165 SAMEA7095588 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095588 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#237 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#237 scientific_name Mus musculus strain C57BL/6J ERS4856166 SAMEA7095589 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095589 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#238 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#238 scientific_name Mus musculus strain C57BL/6J ERS4856167 SAMEA7095590 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095590 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#239 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#239 scientific_name Mus musculus strain C57BL/6J ERS4856168 SAMEA7095591 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095591 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#24 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#24 scientific_name Mus musculus strain C57BL/6J ERS4856169 SAMEA7095592 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095592 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#240 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#240 scientific_name Mus musculus strain C57BL/6J ERS4856170 SAMEA7095593 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095593 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#241 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#241 scientific_name Mus musculus strain C57BL/6J ERS4856171 SAMEA7095594 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095594 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#242 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#242 scientific_name Mus musculus strain C57BL/6J ERS4856172 SAMEA7095595 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095595 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#243 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#243 scientific_name Mus musculus strain C57BL/6J ERS4856173 SAMEA7095596 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095596 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#244 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#244 scientific_name Mus musculus strain C57BL/6J ERS4856174 SAMEA7095597 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095597 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#245 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#245 scientific_name Mus musculus strain C57BL/6J ERS4856175 SAMEA7095598 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095598 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#246 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#246 scientific_name Mus musculus strain C57BL/6J ERS4856176 SAMEA7095599 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095599 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#247 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#247 scientific_name Mus musculus strain C57BL/6J ERS4856177 SAMEA7095600 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095600 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#248 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#248 scientific_name Mus musculus strain C57BL/6J ERS4856178 SAMEA7095601 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095601 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#249 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#249 scientific_name Mus musculus strain C57BL/6J ERS4856179 SAMEA7095602 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095602 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#25 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#25 scientific_name Mus musculus strain C57BL/6J ERS4856180 SAMEA7095603 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095603 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#250 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#250 scientific_name Mus musculus strain C57BL/6J ERS4856181 SAMEA7095604 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095604 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#251 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#251 scientific_name Mus musculus strain C57BL/6J ERS4856182 SAMEA7095605 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095605 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#252 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#252 scientific_name Mus musculus strain C57BL/6J ERS4856183 SAMEA7095606 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095606 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#253 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#253 scientific_name Mus musculus strain C57BL/6J ERS4856184 SAMEA7095607 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095607 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#254 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#254 scientific_name Mus musculus strain C57BL/6J ERS4856185 SAMEA7095608 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095608 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#255 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#255 scientific_name Mus musculus strain C57BL/6J ERS4856186 SAMEA7095609 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095609 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#256 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#256 scientific_name Mus musculus strain C57BL/6J ERS4856187 SAMEA7095610 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095610 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#257 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#257 scientific_name Mus musculus strain C57BL/6J ERS4856188 SAMEA7095611 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:47Z External Id SAMEA7095611 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:47Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#258 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#258 scientific_name Mus musculus strain C57BL/6J ERS4856189 SAMEA7095612 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095612 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#259 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#259 scientific_name Mus musculus strain C57BL/6J ERS4856190 SAMEA7095613 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095613 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#26 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#26 scientific_name Mus musculus strain C57BL/6J ERS4856191 SAMEA7095614 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095614 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#260 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#260 scientific_name Mus musculus strain C57BL/6J ERS4856192 SAMEA7095615 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095615 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#261 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#261 scientific_name Mus musculus strain C57BL/6J ERS4856193 SAMEA7095616 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095616 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#262 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#262 scientific_name Mus musculus strain C57BL/6J ERS4856194 SAMEA7095617 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095617 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#263 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#263 scientific_name Mus musculus strain C57BL/6J ERS4856195 SAMEA7095618 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095618 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#264 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#264 scientific_name Mus musculus strain C57BL/6J ERS4856196 SAMEA7095619 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095619 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#265 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#265 scientific_name Mus musculus strain C57BL/6J ERS4856197 SAMEA7095620 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095620 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#266 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#266 scientific_name Mus musculus strain C57BL/6J ERS4856198 SAMEA7095621 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095621 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#267 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#267 scientific_name Mus musculus strain C57BL/6J ERS4856199 SAMEA7095622 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095622 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#268 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#268 scientific_name Mus musculus strain C57BL/6J ERS4856200 SAMEA7095623 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095623 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#269 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#269 scientific_name Mus musculus strain C57BL/6J ERS4856201 SAMEA7095624 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095624 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#27 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#27 scientific_name Mus musculus strain C57BL/6J ERS4856202 SAMEA7095625 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095625 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#270 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#270 scientific_name Mus musculus strain C57BL/6J ERS4856203 SAMEA7095626 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095626 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#271 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#271 scientific_name Mus musculus strain C57BL/6J ERS4856204 SAMEA7095627 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095627 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#272 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#272 scientific_name Mus musculus strain C57BL/6J ERS4856205 SAMEA7095628 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095628 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#273 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#273 scientific_name Mus musculus strain C57BL/6J ERS4856206 SAMEA7095629 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095629 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#274 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#274 scientific_name Mus musculus strain C57BL/6J ERS4856207 SAMEA7095630 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095630 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#275 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#275 scientific_name Mus musculus strain C57BL/6J ERS4856208 SAMEA7095631 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095631 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#276 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#276 scientific_name Mus musculus strain C57BL/6J ERS4856209 SAMEA7095632 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095632 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#277 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#277 scientific_name Mus musculus strain C57BL/6J ERS4856210 SAMEA7095633 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095633 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#278 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#278 scientific_name Mus musculus strain C57BL/6J ERS4856211 SAMEA7095634 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095634 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#279 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#279 scientific_name Mus musculus strain C57BL/6J ERS4856212 SAMEA7095635 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095635 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#28 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#28 scientific_name Mus musculus strain C57BL/6J ERS4856213 SAMEA7095636 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095636 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#280 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#280 scientific_name Mus musculus strain C57BL/6J ERS4856214 SAMEA7095637 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095637 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#281 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#281 scientific_name Mus musculus strain C57BL/6J ERS4856215 SAMEA7095638 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095638 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#282 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#282 scientific_name Mus musculus strain C57BL/6J ERS4856216 SAMEA7095639 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095639 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#283 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#283 scientific_name Mus musculus strain C57BL/6J ERS4856217 SAMEA7095640 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095640 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#284 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#284 scientific_name Mus musculus strain C57BL/6J ERS4856218 SAMEA7095641 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095641 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#285 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#285 scientific_name Mus musculus strain C57BL/6J ERS4856219 SAMEA7095642 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095642 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#286 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#286 scientific_name Mus musculus strain C57BL/6J ERS4856220 SAMEA7095643 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095643 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#287 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#287 scientific_name Mus musculus strain C57BL/6J ERS4856221 SAMEA7095644 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095644 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#288 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#288 scientific_name Mus musculus strain C57BL/6J ERS4856222 SAMEA7095645 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095645 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#289 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#289 scientific_name Mus musculus strain C57BL/6J ERS4856223 SAMEA7095646 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095646 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#29 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#29 scientific_name Mus musculus strain C57BL/6J ERS4856224 SAMEA7095647 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095647 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#290 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#290 scientific_name Mus musculus strain C57BL/6J ERS4856225 SAMEA7095648 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095648 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#291 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#291 scientific_name Mus musculus strain C57BL/6J ERS4856226 SAMEA7095649 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095649 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#292 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#292 scientific_name Mus musculus strain C57BL/6J ERS4856227 SAMEA7095650 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095650 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#293 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#293 scientific_name Mus musculus strain C57BL/6J ERS4856228 SAMEA7095651 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095651 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#294 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#294 scientific_name Mus musculus strain C57BL/6J ERS4856229 SAMEA7095652 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095652 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#295 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#295 scientific_name Mus musculus strain C57BL/6J ERS4856230 SAMEA7095653 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095653 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#296 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#296 scientific_name Mus musculus strain C57BL/6J ERS4856231 SAMEA7095654 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095654 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#297 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#297 scientific_name Mus musculus strain C57BL/6J ERS4856232 SAMEA7095655 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095655 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#298 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#298 scientific_name Mus musculus strain C57BL/6J ERS4856233 SAMEA7095656 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095656 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#299 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#299 scientific_name Mus musculus strain C57BL/6J ERS4856234 SAMEA7095657 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095657 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#3 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#3 scientific_name Mus musculus strain C57BL/6J ERS4856235 SAMEA7095658 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095658 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#30 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#30 scientific_name Mus musculus strain C57BL/6J ERS4856236 SAMEA7095659 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095659 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#300 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#300 scientific_name Mus musculus strain C57BL/6J ERS4856237 SAMEA7095660 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095660 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#301 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#301 scientific_name Mus musculus strain C57BL/6J ERS4856238 SAMEA7095661 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095661 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#302 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#302 scientific_name Mus musculus strain C57BL/6J ERS4856239 SAMEA7095662 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095662 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#303 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#303 scientific_name Mus musculus strain C57BL/6J ERS4856240 SAMEA7095663 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095663 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#304 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#304 scientific_name Mus musculus strain C57BL/6J ERS4856241 SAMEA7095664 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095664 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#305 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#305 scientific_name Mus musculus strain C57BL/6J ERS4856242 SAMEA7095665 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095665 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#306 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#306 scientific_name Mus musculus strain C57BL/6J ERS4856243 SAMEA7095666 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095666 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#307 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#307 scientific_name Mus musculus strain C57BL/6J ERS4856244 SAMEA7095667 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095667 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#308 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#308 scientific_name Mus musculus strain C57BL/6J ERS4856245 SAMEA7095668 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095668 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#309 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#309 scientific_name Mus musculus strain C57BL/6J ERS4856246 SAMEA7095669 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095669 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#31 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#31 scientific_name Mus musculus strain C57BL/6J ERS4856247 SAMEA7095670 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095670 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#310 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#310 scientific_name Mus musculus strain C57BL/6J ERS4856248 SAMEA7095671 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095671 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#311 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#311 scientific_name Mus musculus strain C57BL/6J ERS4856249 SAMEA7095672 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095672 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#312 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#312 scientific_name Mus musculus strain C57BL/6J ERS4856250 SAMEA7095673 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095673 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#313 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#313 scientific_name Mus musculus strain C57BL/6J ERS4856251 SAMEA7095674 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095674 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#314 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#314 scientific_name Mus musculus strain C57BL/6J ERS4856252 SAMEA7095675 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095675 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#315 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#315 scientific_name Mus musculus strain C57BL/6J ERS4856253 SAMEA7095676 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095676 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#316 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#316 scientific_name Mus musculus strain C57BL/6J ERS4856254 SAMEA7095677 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095677 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#317 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#317 scientific_name Mus musculus strain C57BL/6J ERS4856255 SAMEA7095678 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095678 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#318 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#318 scientific_name Mus musculus strain C57BL/6J ERS4856256 SAMEA7095679 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095679 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#319 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#319 scientific_name Mus musculus strain C57BL/6J ERS4856257 SAMEA7095680 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095680 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#32 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#32 scientific_name Mus musculus strain C57BL/6J ERS4856258 SAMEA7095681 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095681 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#320 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#320 scientific_name Mus musculus strain C57BL/6J ERS4856259 SAMEA7095682 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095682 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#321 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#321 scientific_name Mus musculus strain C57BL/6J ERS4856260 SAMEA7095683 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095683 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#322 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#322 scientific_name Mus musculus strain C57BL/6J ERS4856261 SAMEA7095684 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095684 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#323 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#323 scientific_name Mus musculus strain C57BL/6J ERS4856262 SAMEA7095685 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095685 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#324 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#324 scientific_name Mus musculus strain C57BL/6J ERS4856263 SAMEA7095686 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095686 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#325 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#325 scientific_name Mus musculus strain C57BL/6J ERS4856264 SAMEA7095687 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095687 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#326 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#326 scientific_name Mus musculus strain C57BL/6J ERS4856265 SAMEA7095688 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095688 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#327 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#327 scientific_name Mus musculus strain C57BL/6J ERS4856266 SAMEA7095689 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095689 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#328 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#328 scientific_name Mus musculus strain C57BL/6J ERS4856267 SAMEA7095690 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095690 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#329 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#329 scientific_name Mus musculus strain C57BL/6J ERS4856268 SAMEA7095691 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095691 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#33 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#33 scientific_name Mus musculus strain C57BL/6J ERS4856269 SAMEA7095692 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095692 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#330 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#330 scientific_name Mus musculus strain C57BL/6J ERS4856270 SAMEA7095693 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095693 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#331 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#331 scientific_name Mus musculus strain C57BL/6J ERS4856271 SAMEA7095694 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:48Z External Id SAMEA7095694 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:48Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#332 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#332 scientific_name Mus musculus strain C57BL/6J ERS4856272 SAMEA7095695 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095695 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#333 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#333 scientific_name Mus musculus strain C57BL/6J ERS4856273 SAMEA7095696 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095696 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#334 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#334 scientific_name Mus musculus strain C57BL/6J ERS4856274 SAMEA7095697 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095697 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#335 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#335 scientific_name Mus musculus strain C57BL/6J ERS4856275 SAMEA7095698 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095698 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#336 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#336 scientific_name Mus musculus strain C57BL/6J ERS4856277 SAMEA7095700 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095700 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#338 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#338 scientific_name Mus musculus strain C57BL/6J ERS4856278 SAMEA7095701 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095701 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#339 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#339 scientific_name Mus musculus strain C57BL/6J ERS4856279 SAMEA7095702 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095702 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#34 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#34 scientific_name Mus musculus strain C57BL/6J ERS4856280 SAMEA7095703 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095703 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#340 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#340 scientific_name Mus musculus strain C57BL/6J ERS4856281 SAMEA7095704 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095704 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#341 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#341 scientific_name Mus musculus strain C57BL/6J ERS4856282 SAMEA7095705 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095705 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#342 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#342 scientific_name Mus musculus strain C57BL/6J ERS4856283 SAMEA7095706 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095706 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#343 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#343 scientific_name Mus musculus strain C57BL/6J ERS4856284 SAMEA7095707 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095707 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#344 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#344 scientific_name Mus musculus strain C57BL/6J ERS4856285 SAMEA7095708 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095708 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#345 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#345 scientific_name Mus musculus strain C57BL/6J ERS4856286 SAMEA7095709 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095709 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#346 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#346 scientific_name Mus musculus strain C57BL/6J ERS4856276 SAMEA7095699 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095699 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#337 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#337 scientific_name Mus musculus strain C57BL/6J ERS4856287 SAMEA7095710 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095710 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#347 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#347 scientific_name Mus musculus strain C57BL/6J ERS4856288 SAMEA7095711 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095711 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#348 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#348 scientific_name Mus musculus strain C57BL/6J ERS4856289 SAMEA7095712 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095712 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#349 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#349 scientific_name Mus musculus strain C57BL/6J ERS4856290 SAMEA7095713 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095713 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#35 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#35 scientific_name Mus musculus strain C57BL/6J ERS4856291 SAMEA7095714 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095714 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#350 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#350 scientific_name Mus musculus strain C57BL/6J ERS4856292 SAMEA7095715 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095715 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#351 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#351 scientific_name Mus musculus strain C57BL/6J ERS4856293 SAMEA7095716 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095716 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#352 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#352 scientific_name Mus musculus strain C57BL/6J ERS4856294 SAMEA7095717 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095717 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#353 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#353 scientific_name Mus musculus strain C57BL/6J ERS4856295 SAMEA7095718 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095718 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#354 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#354 scientific_name Mus musculus strain C57BL/6J ERS4856296 SAMEA7095719 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095719 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#355 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#355 scientific_name Mus musculus strain C57BL/6J ERS4856297 SAMEA7095720 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095720 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#356 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#356 scientific_name Mus musculus strain C57BL/6J ERS4856298 SAMEA7095721 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095721 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#357 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#357 scientific_name Mus musculus strain C57BL/6J ERS4856299 SAMEA7095722 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095722 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#358 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#358 scientific_name Mus musculus strain C57BL/6J ERS4856300 SAMEA7095723 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095723 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#359 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#359 scientific_name Mus musculus strain C57BL/6J ERS4856301 SAMEA7095724 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095724 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#36 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#36 scientific_name Mus musculus strain C57BL/6J ERS4856302 SAMEA7095725 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095725 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#360 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#360 scientific_name Mus musculus strain C57BL/6J ERS4856303 SAMEA7095726 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095726 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#361 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#361 scientific_name Mus musculus strain C57BL/6J ERS4856304 SAMEA7095727 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095727 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#362 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#362 scientific_name Mus musculus strain C57BL/6J ERS4856305 SAMEA7095728 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095728 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#363 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#363 scientific_name Mus musculus strain C57BL/6J ERS4856306 SAMEA7095729 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095729 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#364 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#364 scientific_name Mus musculus strain C57BL/6J ERS4856307 SAMEA7095730 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095730 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#365 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#365 scientific_name Mus musculus strain C57BL/6J ERS4856308 SAMEA7095731 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095731 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#366 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#366 scientific_name Mus musculus strain C57BL/6J ERS4856309 SAMEA7095732 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095732 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#367 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#367 scientific_name Mus musculus strain C57BL/6J ERS4856310 SAMEA7095733 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095733 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#368 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#368 scientific_name Mus musculus strain C57BL/6J ERS4856311 SAMEA7095734 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095734 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#369 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#369 scientific_name Mus musculus strain C57BL/6J ERS4856312 SAMEA7095735 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095735 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#37 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#37 scientific_name Mus musculus strain C57BL/6J ERS4856313 SAMEA7095736 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095736 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#370 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#370 scientific_name Mus musculus strain C57BL/6J ERS4856314 SAMEA7095737 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095737 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#371 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#371 scientific_name Mus musculus strain C57BL/6J ERS4856315 SAMEA7095738 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095738 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#372 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_1#372 scientific_name Mus musculus strain C57BL/6J ERS4856316 SAMEA7095739 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095739 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#373 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#373 scientific_name Mus musculus strain C57BL/6J ERS4856317 SAMEA7095740 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095740 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#374 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#374 scientific_name Mus musculus strain C57BL/6J ERS4856318 SAMEA7095741 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095741 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#375 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#375 scientific_name Mus musculus strain C57BL/6J ERS4856319 SAMEA7095742 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095742 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#376 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#376 scientific_name Mus musculus strain C57BL/6J ERS4856320 SAMEA7095743 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095743 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#377 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#377 scientific_name Mus musculus strain C57BL/6J ERS4856321 SAMEA7095744 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095744 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#378 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#378 scientific_name Mus musculus strain C57BL/6J ERS4856322 SAMEA7095745 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095745 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#379 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#379 scientific_name Mus musculus strain C57BL/6J ERS4856323 SAMEA7095746 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095746 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#38 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#38 scientific_name Mus musculus strain C57BL/6J ERS4856324 SAMEA7095747 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095747 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#380 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#380 scientific_name Mus musculus strain C57BL/6J ERS4856325 SAMEA7095748 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095748 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#381 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#381 scientific_name Mus musculus strain C57BL/6J ERS4856326 SAMEA7095749 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095749 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#382 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#382 scientific_name Mus musculus strain C57BL/6J ERS4856327 SAMEA7095750 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095750 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#383 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#383 scientific_name Mus musculus strain C57BL/6J ERS4856328 SAMEA7095751 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095751 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#384 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_1#384 scientific_name Mus musculus strain C57BL/6J ERS4856329 SAMEA7095752 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095752 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#39 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#39 scientific_name Mus musculus strain C57BL/6J ERS4856330 SAMEA7095753 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095753 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#4 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#4 scientific_name Mus musculus strain C57BL/6J ERS4856331 SAMEA7095754 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095754 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#40 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#40 scientific_name Mus musculus strain C57BL/6J ERS4856332 SAMEA7095755 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095755 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#41 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#41 scientific_name Mus musculus strain C57BL/6J ERS4856333 SAMEA7095756 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095756 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#42 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#42 scientific_name Mus musculus strain C57BL/6J ERS4856334 SAMEA7095757 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095757 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#43 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#43 scientific_name Mus musculus strain C57BL/6J ERS4856335 SAMEA7095758 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095758 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#44 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#44 scientific_name Mus musculus strain C57BL/6J ERS4856336 SAMEA7095759 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095759 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#45 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#45 scientific_name Mus musculus strain C57BL/6J ERS4856337 SAMEA7095760 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095760 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#46 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#46 scientific_name Mus musculus strain C57BL/6J ERS4856338 SAMEA7095761 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095761 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#47 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#47 scientific_name Mus musculus strain C57BL/6J ERS4856339 SAMEA7095762 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095762 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#48 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#48 scientific_name Mus musculus strain C57BL/6J ERS4856340 SAMEA7095763 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095763 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#49 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#49 scientific_name Mus musculus strain C57BL/6J ERS4856341 SAMEA7095764 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095764 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#5 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#5 scientific_name Mus musculus strain C57BL/6J ERS4856342 SAMEA7095765 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095765 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#50 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#50 scientific_name Mus musculus strain C57BL/6J ERS4856343 SAMEA7095766 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095766 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#51 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#51 scientific_name Mus musculus strain C57BL/6J ERS4856344 SAMEA7095767 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095767 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#52 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#52 scientific_name Mus musculus strain C57BL/6J ERS4856345 SAMEA7095768 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095768 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#53 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#53 scientific_name Mus musculus strain C57BL/6J ERS4856346 SAMEA7095769 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095769 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#54 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#54 scientific_name Mus musculus strain C57BL/6J ERS4856347 SAMEA7095770 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095770 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#55 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#55 scientific_name Mus musculus strain C57BL/6J ERS4856348 SAMEA7095771 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095771 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#56 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#56 scientific_name Mus musculus strain C57BL/6J ERS4856349 SAMEA7095772 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095772 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#57 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#57 scientific_name Mus musculus strain C57BL/6J ERS4856350 SAMEA7095773 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095773 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#58 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#58 scientific_name Mus musculus strain C57BL/6J ERS4856351 SAMEA7095774 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095774 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#59 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#59 scientific_name Mus musculus strain C57BL/6J ERS4856352 SAMEA7095775 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095775 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#6 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#6 scientific_name Mus musculus strain C57BL/6J ERS4856353 SAMEA7095776 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095776 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#60 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#60 scientific_name Mus musculus strain C57BL/6J ERS4856354 SAMEA7095777 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095777 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#61 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#61 scientific_name Mus musculus strain C57BL/6J ERS4856355 SAMEA7095778 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:49Z External Id SAMEA7095778 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:49Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#62 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#62 scientific_name Mus musculus strain C57BL/6J ERS4856356 SAMEA7095779 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095779 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#63 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#63 scientific_name Mus musculus strain C57BL/6J ERS4856357 SAMEA7095780 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095780 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#64 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#64 scientific_name Mus musculus strain C57BL/6J ERS4856358 SAMEA7095781 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095781 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#65 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#65 scientific_name Mus musculus strain C57BL/6J ERS4856359 SAMEA7095782 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095782 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#66 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#66 scientific_name Mus musculus strain C57BL/6J ERS4856360 SAMEA7095783 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095783 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#67 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#67 scientific_name Mus musculus strain C57BL/6J ERS4856361 SAMEA7095784 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095784 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#68 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#68 scientific_name Mus musculus strain C57BL/6J ERS4856362 SAMEA7095785 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095785 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#69 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#69 scientific_name Mus musculus strain C57BL/6J ERS4856363 SAMEA7095786 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095786 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#7 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#7 scientific_name Mus musculus strain C57BL/6J ERS4856364 SAMEA7095787 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095787 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#70 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#70 scientific_name Mus musculus strain C57BL/6J ERS4856365 SAMEA7095788 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095788 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#71 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#71 scientific_name Mus musculus strain C57BL/6J ERS4856366 SAMEA7095789 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095789 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#72 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#72 scientific_name Mus musculus strain C57BL/6J ERS4856367 SAMEA7095790 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095790 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#73 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#73 scientific_name Mus musculus strain C57BL/6J ERS4856368 SAMEA7095791 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095791 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#74 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#74 scientific_name Mus musculus strain C57BL/6J ERS4856369 SAMEA7095792 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095792 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#75 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#75 scientific_name Mus musculus strain C57BL/6J ERS4856370 SAMEA7095793 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095793 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#76 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#76 scientific_name Mus musculus strain C57BL/6J ERS4856371 SAMEA7095794 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095794 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#77 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#77 scientific_name Mus musculus strain C57BL/6J ERS4856372 SAMEA7095795 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095795 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#78 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#78 scientific_name Mus musculus strain C57BL/6J ERS4856373 SAMEA7095796 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095796 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#79 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#79 scientific_name Mus musculus strain C57BL/6J ERS4856374 SAMEA7095797 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095797 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#8 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#8 scientific_name Mus musculus strain C57BL/6J ERS4856375 SAMEA7095798 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095798 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#80 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#80 scientific_name Mus musculus strain C57BL/6J ERS4856376 SAMEA7095799 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095799 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#81 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#81 scientific_name Mus musculus strain C57BL/6J ERS4856377 SAMEA7095800 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095800 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#82 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#82 scientific_name Mus musculus strain C57BL/6J ERS4856378 SAMEA7095801 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095801 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#83 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#83 scientific_name Mus musculus strain C57BL/6J ERS4856379 SAMEA7095802 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095802 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#84 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#84 scientific_name Mus musculus strain C57BL/6J ERS4856380 SAMEA7095803 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095803 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#85 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#85 scientific_name Mus musculus strain C57BL/6J ERS4856381 SAMEA7095804 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095804 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#86 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#86 scientific_name Mus musculus strain C57BL/6J ERS4856382 SAMEA7095805 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095805 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#87 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#87 scientific_name Mus musculus strain C57BL/6J ERS4856383 SAMEA7095806 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095806 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#88 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#88 scientific_name Mus musculus strain C57BL/6J ERS4856384 SAMEA7095807 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095807 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#89 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#89 scientific_name Mus musculus strain C57BL/6J ERS4856385 SAMEA7095808 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095808 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#9 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#9 scientific_name Mus musculus strain C57BL/6J ERS4856386 SAMEA7095809 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095809 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#90 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#90 scientific_name Mus musculus strain C57BL/6J ERS4856387 SAMEA7095810 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095810 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#91 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#91 scientific_name Mus musculus strain C57BL/6J ERS4856388 SAMEA7095811 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095811 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#92 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#92 scientific_name Mus musculus strain C57BL/6J ERS4856389 SAMEA7095812 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095812 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#93 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#93 scientific_name Mus musculus strain C57BL/6J ERS4856390 SAMEA7095813 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095813 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#94 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#94 scientific_name Mus musculus strain C57BL/6J ERS4856391 SAMEA7095814 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095814 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#95 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#95 scientific_name Mus musculus strain C57BL/6J ERS4856392 SAMEA7095815 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095815 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#96 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_1#96 scientific_name Mus musculus strain C57BL/6J ERS4856393 SAMEA7095816 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095816 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#97 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#97 scientific_name Mus musculus strain C57BL/6J ERS4856394 SAMEA7095817 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095817 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#98 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#98 scientific_name Mus musculus strain C57BL/6J ERS4856395 SAMEA7095818 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095818 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_1#99 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_1#99 scientific_name Mus musculus strain C57BL/6J ERS4856396 SAMEA7095819 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095819 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#1 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#1 scientific_name Mus musculus strain C57BL/6J ERS4856397 SAMEA7095820 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095820 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#10 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#10 scientific_name Mus musculus strain C57BL/6J ERS4856398 SAMEA7095821 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095821 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#100 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#100 scientific_name Mus musculus strain C57BL/6J ERS4856399 SAMEA7095822 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095822 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#101 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#101 scientific_name Mus musculus strain C57BL/6J ERS4856400 SAMEA7095823 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095823 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#102 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#102 scientific_name Mus musculus strain C57BL/6J ERS4856401 SAMEA7095824 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095824 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#103 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#103 scientific_name Mus musculus strain C57BL/6J ERS4856402 SAMEA7095825 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095825 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#104 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#104 scientific_name Mus musculus strain C57BL/6J ERS4856403 SAMEA7095826 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095826 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#105 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#105 scientific_name Mus musculus strain C57BL/6J ERS4856404 SAMEA7095827 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095827 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#106 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#106 scientific_name Mus musculus strain C57BL/6J ERS4856405 SAMEA7095828 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095828 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#107 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#107 scientific_name Mus musculus strain C57BL/6J ERS4856406 SAMEA7095829 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095829 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#108 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#108 scientific_name Mus musculus strain C57BL/6J ERS4856407 SAMEA7095830 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095830 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#109 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#109 scientific_name Mus musculus strain C57BL/6J ERS4856408 SAMEA7095831 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095831 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#11 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#11 scientific_name Mus musculus strain C57BL/6J ERS4856409 SAMEA7095832 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095832 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#110 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#110 scientific_name Mus musculus strain C57BL/6J ERS4856410 SAMEA7095833 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095833 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#111 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#111 scientific_name Mus musculus strain C57BL/6J ERS4856411 SAMEA7095834 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095834 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#112 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#112 scientific_name Mus musculus strain C57BL/6J ERS4856412 SAMEA7095835 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095835 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#113 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#113 scientific_name Mus musculus strain C57BL/6J ERS4856413 SAMEA7095836 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095836 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#114 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#114 scientific_name Mus musculus strain C57BL/6J ERS4856414 SAMEA7095837 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095837 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#115 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#115 scientific_name Mus musculus strain C57BL/6J ERS4856415 SAMEA7095838 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095838 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#116 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#116 scientific_name Mus musculus strain C57BL/6J ERS4856416 SAMEA7095839 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095839 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#117 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#117 scientific_name Mus musculus strain C57BL/6J ERS4856417 SAMEA7095840 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095840 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#118 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#118 scientific_name Mus musculus strain C57BL/6J ERS4856418 SAMEA7095841 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095841 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#119 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#119 scientific_name Mus musculus strain C57BL/6J ERS4856419 SAMEA7095842 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095842 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#12 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#12 scientific_name Mus musculus strain C57BL/6J ERS4856420 SAMEA7095843 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095843 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#120 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#120 scientific_name Mus musculus strain C57BL/6J ERS4856421 SAMEA7095844 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095844 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#121 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#121 scientific_name Mus musculus strain C57BL/6J ERS4856422 SAMEA7095845 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095845 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#122 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#122 scientific_name Mus musculus strain C57BL/6J ERS4856423 SAMEA7095846 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095846 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#123 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#123 scientific_name Mus musculus strain C57BL/6J ERS4856424 SAMEA7095847 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095847 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#124 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#124 scientific_name Mus musculus strain C57BL/6J ERS4856425 SAMEA7095848 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095848 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#125 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#125 scientific_name Mus musculus strain C57BL/6J ERS4856426 SAMEA7095849 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095849 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#126 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#126 scientific_name Mus musculus strain C57BL/6J ERS4856427 SAMEA7095850 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095850 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#127 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#127 scientific_name Mus musculus strain C57BL/6J ERS4856428 SAMEA7095851 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095851 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#128 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#128 scientific_name Mus musculus strain C57BL/6J ERS4856429 SAMEA7095852 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095852 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#129 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#129 scientific_name Mus musculus strain C57BL/6J ERS4856430 SAMEA7095853 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095853 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#13 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#13 scientific_name Mus musculus strain C57BL/6J ERS4856431 SAMEA7095854 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095854 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#130 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#130 scientific_name Mus musculus strain C57BL/6J ERS4856432 SAMEA7095855 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095855 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#131 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#131 scientific_name Mus musculus strain C57BL/6J ERS4856433 SAMEA7095856 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095856 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#132 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#132 scientific_name Mus musculus strain C57BL/6J ERS4856434 SAMEA7095857 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095857 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#133 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#133 scientific_name Mus musculus strain C57BL/6J ERS4856435 SAMEA7095858 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095858 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#134 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#134 scientific_name Mus musculus strain C57BL/6J ERS4856436 SAMEA7095859 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095859 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#135 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#135 scientific_name Mus musculus strain C57BL/6J ERS4856437 SAMEA7095860 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:50Z External Id SAMEA7095860 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:50Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#136 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#136 scientific_name Mus musculus strain C57BL/6J ERS4856438 SAMEA7095861 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095861 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#137 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#137 scientific_name Mus musculus strain C57BL/6J ERS4856439 SAMEA7095862 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095862 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#138 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#138 scientific_name Mus musculus strain C57BL/6J ERS4856440 SAMEA7095863 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095863 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#139 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#139 scientific_name Mus musculus strain C57BL/6J ERS4856441 SAMEA7095864 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095864 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#14 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#14 scientific_name Mus musculus strain C57BL/6J ERS4856442 SAMEA7095865 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095865 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#140 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#140 scientific_name Mus musculus strain C57BL/6J ERS4856443 SAMEA7095866 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095866 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#141 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#141 scientific_name Mus musculus strain C57BL/6J ERS4856444 SAMEA7095867 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095867 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#142 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#142 scientific_name Mus musculus strain C57BL/6J ERS4856445 SAMEA7095868 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095868 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#143 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#143 scientific_name Mus musculus strain C57BL/6J ERS4856446 SAMEA7095869 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095869 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#144 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#144 scientific_name Mus musculus strain C57BL/6J ERS4856447 SAMEA7095870 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095870 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#145 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#145 scientific_name Mus musculus strain C57BL/6J ERS4856448 SAMEA7095871 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095871 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#146 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#146 scientific_name Mus musculus strain C57BL/6J ERS4856449 SAMEA7095872 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095872 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#147 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#147 scientific_name Mus musculus strain C57BL/6J ERS4856450 SAMEA7095873 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095873 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#148 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#148 scientific_name Mus musculus strain C57BL/6J ERS4856451 SAMEA7095874 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095874 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#149 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#149 scientific_name Mus musculus strain C57BL/6J ERS4856452 SAMEA7095875 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095875 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#15 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#15 scientific_name Mus musculus strain C57BL/6J ERS4856453 SAMEA7095876 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095876 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#150 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#150 scientific_name Mus musculus strain C57BL/6J ERS4856454 SAMEA7095877 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095877 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#151 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#151 scientific_name Mus musculus strain C57BL/6J ERS4856455 SAMEA7095878 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095878 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#152 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#152 scientific_name Mus musculus strain C57BL/6J ERS4856456 SAMEA7095879 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095879 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#153 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#153 scientific_name Mus musculus strain C57BL/6J ERS4856457 SAMEA7095880 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095880 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#154 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#154 scientific_name Mus musculus strain C57BL/6J ERS4856458 SAMEA7095881 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095881 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#155 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#155 scientific_name Mus musculus strain C57BL/6J ERS4856459 SAMEA7095882 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095882 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#156 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#156 scientific_name Mus musculus strain C57BL/6J ERS4856460 SAMEA7095883 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095883 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#157 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#157 scientific_name Mus musculus strain C57BL/6J ERS4856461 SAMEA7095884 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095884 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#158 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#158 scientific_name Mus musculus strain C57BL/6J ERS4856462 SAMEA7095885 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095885 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#159 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#159 scientific_name Mus musculus strain C57BL/6J ERS4856463 SAMEA7095886 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095886 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#16 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#16 scientific_name Mus musculus strain C57BL/6J ERS4856464 SAMEA7095887 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095887 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#160 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#160 scientific_name Mus musculus strain C57BL/6J ERS4856465 SAMEA7095888 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095888 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#161 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#161 scientific_name Mus musculus strain C57BL/6J ERS4856466 SAMEA7095889 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095889 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#162 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#162 scientific_name Mus musculus strain C57BL/6J ERS4856467 SAMEA7095890 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095890 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#163 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#163 scientific_name Mus musculus strain C57BL/6J ERS4856468 SAMEA7095891 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095891 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#164 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#164 scientific_name Mus musculus strain C57BL/6J ERS4856469 SAMEA7095892 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095892 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#165 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#165 scientific_name Mus musculus strain C57BL/6J ERS4856470 SAMEA7095893 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095893 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#166 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#166 scientific_name Mus musculus strain C57BL/6J ERS4856471 SAMEA7095894 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095894 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#167 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#167 scientific_name Mus musculus strain C57BL/6J ERS4856472 SAMEA7095895 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095895 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#169 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#169 scientific_name Mus musculus strain C57BL/6J ERS4856473 SAMEA7095896 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095896 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#17 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#17 scientific_name Mus musculus strain C57BL/6J ERS4856474 SAMEA7095897 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095897 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#170 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#170 scientific_name Mus musculus strain C57BL/6J ERS4856475 SAMEA7095898 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095898 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#171 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#171 scientific_name Mus musculus strain C57BL/6J ERS4856476 SAMEA7095899 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095899 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#172 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#172 scientific_name Mus musculus strain C57BL/6J ERS4856477 SAMEA7095900 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095900 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#173 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#173 scientific_name Mus musculus strain C57BL/6J ERS4856478 SAMEA7095901 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095901 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#174 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#174 scientific_name Mus musculus strain C57BL/6J ERS4856479 SAMEA7095902 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095902 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#175 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#175 scientific_name Mus musculus strain C57BL/6J ERS4856480 SAMEA7095903 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095903 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#176 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#176 scientific_name Mus musculus strain C57BL/6J ERS4856481 SAMEA7095904 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095904 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#177 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#177 scientific_name Mus musculus strain C57BL/6J ERS4856482 SAMEA7095905 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095905 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#178 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#178 scientific_name Mus musculus strain C57BL/6J ERS4856483 SAMEA7095906 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095906 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#179 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#179 scientific_name Mus musculus strain C57BL/6J ERS4856484 SAMEA7095907 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095907 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#18 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#18 scientific_name Mus musculus strain C57BL/6J ERS4856485 SAMEA7095908 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095908 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#180 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#180 scientific_name Mus musculus strain C57BL/6J ERS4856486 SAMEA7095909 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095909 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#181 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#181 scientific_name Mus musculus strain C57BL/6J ERS4856487 SAMEA7095910 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095910 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#182 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#182 scientific_name Mus musculus strain C57BL/6J ERS4856488 SAMEA7095911 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095911 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#183 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#183 scientific_name Mus musculus strain C57BL/6J ERS4856489 SAMEA7095912 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095912 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#184 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#184 scientific_name Mus musculus strain C57BL/6J ERS4856490 SAMEA7095913 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095913 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#185 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#185 scientific_name Mus musculus strain C57BL/6J ERS4856491 SAMEA7095914 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095914 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#186 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#186 scientific_name Mus musculus strain C57BL/6J ERS4856492 SAMEA7095915 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095915 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#187 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#187 scientific_name Mus musculus strain C57BL/6J ERS4856493 SAMEA7095916 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095916 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#188 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#188 scientific_name Mus musculus strain C57BL/6J ERS4856494 SAMEA7095917 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095917 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#189 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#189 scientific_name Mus musculus strain C57BL/6J ERS4856495 SAMEA7095918 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095918 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#19 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#19 scientific_name Mus musculus strain C57BL/6J ERS4856496 SAMEA7095919 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095919 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#190 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#190 scientific_name Mus musculus strain C57BL/6J ERS4856497 SAMEA7095920 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095920 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#191 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#191 scientific_name Mus musculus strain C57BL/6J ERS4856498 SAMEA7095921 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095921 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#192 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#192 scientific_name Mus musculus strain C57BL/6J ERS4856499 SAMEA7095922 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095922 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#193 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#193 scientific_name Mus musculus strain C57BL/6J ERS4856500 SAMEA7095923 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095923 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#194 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#194 scientific_name Mus musculus strain C57BL/6J ERS4856501 SAMEA7095924 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095924 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#195 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#195 scientific_name Mus musculus strain C57BL/6J ERS4856502 SAMEA7095925 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095925 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#196 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#196 scientific_name Mus musculus strain C57BL/6J ERS4856503 SAMEA7095926 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095926 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#197 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#197 scientific_name Mus musculus strain C57BL/6J ERS4856504 SAMEA7095927 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095927 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#198 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#198 scientific_name Mus musculus strain C57BL/6J ERS4856505 SAMEA7095928 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095928 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#199 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#199 scientific_name Mus musculus strain C57BL/6J ERS4856506 SAMEA7095929 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095929 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#2 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#2 scientific_name Mus musculus strain C57BL/6J ERS4856507 SAMEA7095930 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095930 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#20 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#20 scientific_name Mus musculus strain C57BL/6J ERS4856508 SAMEA7095931 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095931 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#200 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#200 scientific_name Mus musculus strain C57BL/6J ERS4856509 SAMEA7095932 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095932 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#201 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#201 scientific_name Mus musculus strain C57BL/6J ERS4856510 SAMEA7095933 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095933 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#202 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#202 scientific_name Mus musculus strain C57BL/6J ERS4856511 SAMEA7095934 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095934 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#203 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#203 scientific_name Mus musculus strain C57BL/6J ERS4856512 SAMEA7095935 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095935 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#204 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#204 scientific_name Mus musculus strain C57BL/6J ERS4856513 SAMEA7095936 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095936 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#205 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#205 scientific_name Mus musculus strain C57BL/6J ERS4856514 SAMEA7095937 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095937 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#206 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#206 scientific_name Mus musculus strain C57BL/6J ERS4856515 SAMEA7095938 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095938 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#207 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#207 scientific_name Mus musculus strain C57BL/6J ERS4856516 SAMEA7095939 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095939 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#208 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#208 scientific_name Mus musculus strain C57BL/6J ERS4856517 SAMEA7095940 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095940 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#209 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#209 scientific_name Mus musculus strain C57BL/6J ERS4856518 SAMEA7095941 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:51Z External Id SAMEA7095941 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:51Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#21 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#21 scientific_name Mus musculus strain C57BL/6J ERS4856519 SAMEA7095942 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095942 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#210 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#210 scientific_name Mus musculus strain C57BL/6J ERS4856520 SAMEA7095943 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095943 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#211 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#211 scientific_name Mus musculus strain C57BL/6J ERS4856521 SAMEA7095944 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095944 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#212 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#212 scientific_name Mus musculus strain C57BL/6J ERS4856522 SAMEA7095945 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095945 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#213 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#213 scientific_name Mus musculus strain C57BL/6J ERS4856523 SAMEA7095946 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095946 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#214 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#214 scientific_name Mus musculus strain C57BL/6J ERS4856524 SAMEA7095947 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095947 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#215 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#215 scientific_name Mus musculus strain C57BL/6J ERS4856525 SAMEA7095948 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095948 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#216 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#216 scientific_name Mus musculus strain C57BL/6J ERS4856526 SAMEA7095949 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095949 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#217 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#217 scientific_name Mus musculus strain C57BL/6J ERS4856527 SAMEA7095950 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095950 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#218 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#218 scientific_name Mus musculus strain C57BL/6J ERS4856528 SAMEA7095951 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095951 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#219 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#219 scientific_name Mus musculus strain C57BL/6J ERS4856529 SAMEA7095952 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095952 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#22 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#22 scientific_name Mus musculus strain C57BL/6J ERS4856530 SAMEA7095953 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095953 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#220 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#220 scientific_name Mus musculus strain C57BL/6J ERS4856531 SAMEA7095954 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095954 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#221 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#221 scientific_name Mus musculus strain C57BL/6J ERS4856532 SAMEA7095955 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095955 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#222 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#222 scientific_name Mus musculus strain C57BL/6J ERS4856533 SAMEA7095956 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095956 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#223 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#223 scientific_name Mus musculus strain C57BL/6J ERS4856534 SAMEA7095957 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095957 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#224 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#224 scientific_name Mus musculus strain C57BL/6J ERS4856535 SAMEA7095958 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095958 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#225 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#225 scientific_name Mus musculus strain C57BL/6J ERS4856536 SAMEA7095959 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095959 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#226 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#226 scientific_name Mus musculus strain C57BL/6J ERS4856537 SAMEA7095960 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095960 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#227 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#227 scientific_name Mus musculus strain C57BL/6J ERS4856538 SAMEA7095961 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095961 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#228 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#228 scientific_name Mus musculus strain C57BL/6J ERS4856539 SAMEA7095962 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095962 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#229 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#229 scientific_name Mus musculus strain C57BL/6J ERS4856540 SAMEA7095963 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095963 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#23 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#23 scientific_name Mus musculus strain C57BL/6J ERS4856541 SAMEA7095964 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095964 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#230 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#230 scientific_name Mus musculus strain C57BL/6J ERS4856542 SAMEA7095965 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095965 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#231 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#231 scientific_name Mus musculus strain C57BL/6J ERS4856543 SAMEA7095966 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095966 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#232 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#232 scientific_name Mus musculus strain C57BL/6J ERS4856544 SAMEA7095967 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095967 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#233 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#233 scientific_name Mus musculus strain C57BL/6J ERS4856545 SAMEA7095968 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095968 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#234 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#234 scientific_name Mus musculus strain C57BL/6J ERS4856546 SAMEA7095969 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095969 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#235 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#235 scientific_name Mus musculus strain C57BL/6J ERS4856547 SAMEA7095970 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095970 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#236 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#236 scientific_name Mus musculus strain C57BL/6J ERS4856548 SAMEA7095971 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095971 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#237 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#237 scientific_name Mus musculus strain C57BL/6J ERS4856549 SAMEA7095972 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095972 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#238 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#238 scientific_name Mus musculus strain C57BL/6J ERS4856550 SAMEA7095973 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095973 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#239 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#239 scientific_name Mus musculus strain C57BL/6J ERS4856551 SAMEA7095974 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095974 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#24 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#24 scientific_name Mus musculus strain C57BL/6J ERS4856552 SAMEA7095975 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095975 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#240 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#240 scientific_name Mus musculus strain C57BL/6J ERS4856553 SAMEA7095976 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095976 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#241 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#241 scientific_name Mus musculus strain C57BL/6J ERS4856554 SAMEA7095977 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095977 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#242 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#242 scientific_name Mus musculus strain C57BL/6J ERS4856555 SAMEA7095978 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095978 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#243 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#243 scientific_name Mus musculus strain C57BL/6J ERS4856556 SAMEA7095979 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095979 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#244 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#244 scientific_name Mus musculus strain C57BL/6J ERS4856557 SAMEA7095980 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095980 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#245 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#245 scientific_name Mus musculus strain C57BL/6J ERS4856558 SAMEA7095981 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095981 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#246 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#246 scientific_name Mus musculus strain C57BL/6J ERS4856559 SAMEA7095982 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095982 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#247 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#247 scientific_name Mus musculus strain C57BL/6J ERS4856560 SAMEA7095983 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095983 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#248 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#248 scientific_name Mus musculus strain C57BL/6J ERS4856561 SAMEA7095984 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095984 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#249 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#249 scientific_name Mus musculus strain C57BL/6J ERS4856562 SAMEA7095985 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095985 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#25 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#25 scientific_name Mus musculus strain C57BL/6J ERS4856563 SAMEA7095986 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095986 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#250 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#250 scientific_name Mus musculus strain C57BL/6J ERS4856564 SAMEA7095987 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095987 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#251 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#251 scientific_name Mus musculus strain C57BL/6J ERS4856565 SAMEA7095988 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095988 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#252 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#252 scientific_name Mus musculus strain C57BL/6J ERS4856566 SAMEA7095989 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095989 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#253 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#253 scientific_name Mus musculus strain C57BL/6J ERS4856567 SAMEA7095990 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095990 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#254 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#254 scientific_name Mus musculus strain C57BL/6J ERS4856568 SAMEA7095991 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095991 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#255 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#255 scientific_name Mus musculus strain C57BL/6J ERS4856569 SAMEA7095992 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095992 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#256 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#256 scientific_name Mus musculus strain C57BL/6J ERS4856570 SAMEA7095993 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095993 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#257 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#257 scientific_name Mus musculus strain C57BL/6J ERS4856571 SAMEA7095994 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095994 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#258 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#258 scientific_name Mus musculus strain C57BL/6J ERS4856572 SAMEA7095995 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095995 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#259 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#259 scientific_name Mus musculus strain C57BL/6J ERS4856573 SAMEA7095996 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095996 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#26 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#26 scientific_name Mus musculus strain C57BL/6J ERS4856574 SAMEA7095997 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095997 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#260 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#260 scientific_name Mus musculus strain C57BL/6J ERS4856575 SAMEA7095998 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095998 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#261 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#261 scientific_name Mus musculus strain C57BL/6J ERS4856576 SAMEA7095999 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7095999 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#262 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#262 scientific_name Mus musculus strain C57BL/6J ERS4856577 SAMEA7096000 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096000 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#263 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#263 scientific_name Mus musculus strain C57BL/6J ERS4856578 SAMEA7096001 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096001 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#264 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#264 scientific_name Mus musculus strain C57BL/6J ERS4856579 SAMEA7096002 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096002 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#265 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#265 scientific_name Mus musculus strain C57BL/6J ERS4856580 SAMEA7096003 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096003 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#266 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#266 scientific_name Mus musculus strain C57BL/6J ERS4856581 SAMEA7096004 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096004 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#267 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#267 scientific_name Mus musculus strain C57BL/6J ERS4856582 SAMEA7096005 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096005 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#268 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#268 scientific_name Mus musculus strain C57BL/6J ERS4856583 SAMEA7096006 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096006 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#269 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#269 scientific_name Mus musculus strain C57BL/6J ERS4856584 SAMEA7096007 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096007 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#27 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#27 scientific_name Mus musculus strain C57BL/6J ERS4856585 SAMEA7096008 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096008 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#270 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#270 scientific_name Mus musculus strain C57BL/6J ERS4856586 SAMEA7096009 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096009 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#271 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#271 scientific_name Mus musculus strain C57BL/6J ERS4856587 SAMEA7096010 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096010 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#272 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#272 scientific_name Mus musculus strain C57BL/6J ERS4856588 SAMEA7096011 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096011 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#273 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#273 scientific_name Mus musculus strain C57BL/6J ERS4856589 SAMEA7096012 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096012 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#274 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#274 scientific_name Mus musculus strain C57BL/6J ERS4856590 SAMEA7096013 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096013 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#275 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#275 scientific_name Mus musculus strain C57BL/6J ERS4856591 SAMEA7096014 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096014 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#276 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#276 scientific_name Mus musculus strain C57BL/6J ERS4856592 SAMEA7096015 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096015 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#277 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#277 scientific_name Mus musculus strain C57BL/6J ERS4856593 SAMEA7096016 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096016 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#278 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#278 scientific_name Mus musculus strain C57BL/6J ERS4856594 SAMEA7096017 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096017 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#279 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#279 scientific_name Mus musculus strain C57BL/6J ERS4856595 SAMEA7096018 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096018 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#28 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#28 scientific_name Mus musculus strain C57BL/6J ERS4856596 SAMEA7096019 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096019 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#280 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#280 scientific_name Mus musculus strain C57BL/6J ERS4856597 SAMEA7096020 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096020 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#281 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#281 scientific_name Mus musculus strain C57BL/6J ERS4856598 SAMEA7096021 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096021 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#282 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#282 scientific_name Mus musculus strain C57BL/6J ERS4856599 SAMEA7096022 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096022 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#283 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#283 scientific_name Mus musculus strain C57BL/6J ERS4856600 SAMEA7096023 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096023 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#284 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#284 scientific_name Mus musculus strain C57BL/6J ERS4856601 SAMEA7096024 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:52Z External Id SAMEA7096024 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:52Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#285 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#285 scientific_name Mus musculus strain C57BL/6J ERS4856602 SAMEA7096025 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096025 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#286 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#286 scientific_name Mus musculus strain C57BL/6J ERS4856603 SAMEA7096026 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096026 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#287 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#287 scientific_name Mus musculus strain C57BL/6J ERS4856604 SAMEA7096027 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096027 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#288 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#288 scientific_name Mus musculus strain C57BL/6J ERS4856605 SAMEA7096028 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096028 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#289 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#289 scientific_name Mus musculus strain C57BL/6J ERS4856606 SAMEA7096029 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096029 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#29 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#29 scientific_name Mus musculus strain C57BL/6J ERS4856607 SAMEA7096030 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096030 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#290 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#290 scientific_name Mus musculus strain C57BL/6J ERS4856608 SAMEA7096031 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096031 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#291 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#291 scientific_name Mus musculus strain C57BL/6J ERS4856609 SAMEA7096032 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096032 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#292 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#292 scientific_name Mus musculus strain C57BL/6J ERS4856610 SAMEA7096033 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096033 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#293 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#293 scientific_name Mus musculus strain C57BL/6J ERS4856611 SAMEA7096034 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096034 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#294 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#294 scientific_name Mus musculus strain C57BL/6J ERS4856612 SAMEA7096035 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096035 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#295 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#295 scientific_name Mus musculus strain C57BL/6J ERS4856613 SAMEA7096036 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096036 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#296 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#296 scientific_name Mus musculus strain C57BL/6J ERS4856614 SAMEA7096037 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096037 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#297 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#297 scientific_name Mus musculus strain C57BL/6J ERS4856615 SAMEA7096038 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096038 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#298 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#298 scientific_name Mus musculus strain C57BL/6J ERS4856616 SAMEA7096039 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096039 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#299 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#299 scientific_name Mus musculus strain C57BL/6J ERS4856617 SAMEA7096040 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096040 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#3 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#3 scientific_name Mus musculus strain C57BL/6J ERS4856618 SAMEA7096041 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096041 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#30 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#30 scientific_name Mus musculus strain C57BL/6J ERS4856619 SAMEA7096042 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096042 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#300 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#300 scientific_name Mus musculus strain C57BL/6J ERS4856620 SAMEA7096043 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096043 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#301 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#301 scientific_name Mus musculus strain C57BL/6J ERS4856621 SAMEA7096044 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096044 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#302 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#302 scientific_name Mus musculus strain C57BL/6J ERS4856622 SAMEA7096045 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096045 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#303 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#303 scientific_name Mus musculus strain C57BL/6J ERS4856623 SAMEA7096046 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096046 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#304 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#304 scientific_name Mus musculus strain C57BL/6J ERS4856624 SAMEA7096047 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096047 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#305 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#305 scientific_name Mus musculus strain C57BL/6J ERS4856625 SAMEA7096048 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096048 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#306 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#306 scientific_name Mus musculus strain C57BL/6J ERS4856626 SAMEA7096049 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096049 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#307 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#307 scientific_name Mus musculus strain C57BL/6J ERS4856627 SAMEA7096050 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096050 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#308 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#308 scientific_name Mus musculus strain C57BL/6J ERS4856628 SAMEA7096051 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096051 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#309 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#309 scientific_name Mus musculus strain C57BL/6J ERS4856629 SAMEA7096052 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096052 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#31 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#31 scientific_name Mus musculus strain C57BL/6J ERS4856630 SAMEA7096053 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096053 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#310 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#310 scientific_name Mus musculus strain C57BL/6J ERS4856631 SAMEA7096054 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096054 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#311 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#311 scientific_name Mus musculus strain C57BL/6J ERS4856632 SAMEA7096055 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096055 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#312 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#312 scientific_name Mus musculus strain C57BL/6J ERS4856633 SAMEA7096056 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096056 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#313 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#313 scientific_name Mus musculus strain C57BL/6J ERS4856634 SAMEA7096057 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096057 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#314 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#314 scientific_name Mus musculus strain C57BL/6J ERS4856635 SAMEA7096058 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096058 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#315 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#315 scientific_name Mus musculus strain C57BL/6J ERS4856636 SAMEA7096059 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096059 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#316 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#316 scientific_name Mus musculus strain C57BL/6J ERS4856637 SAMEA7096060 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096060 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#317 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#317 scientific_name Mus musculus strain C57BL/6J ERS4856638 SAMEA7096061 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096061 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#318 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#318 scientific_name Mus musculus strain C57BL/6J ERS4856639 SAMEA7096062 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096062 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#319 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#319 scientific_name Mus musculus strain C57BL/6J ERS4856640 SAMEA7096063 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096063 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#32 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#32 scientific_name Mus musculus strain C57BL/6J ERS4856641 SAMEA7096064 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096064 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#320 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#320 scientific_name Mus musculus strain C57BL/6J ERS4856642 SAMEA7096065 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096065 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#321 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#321 scientific_name Mus musculus strain C57BL/6J ERS4856643 SAMEA7096066 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096066 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#322 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#322 scientific_name Mus musculus strain C57BL/6J ERS4856644 SAMEA7096067 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096067 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#323 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#323 scientific_name Mus musculus strain C57BL/6J ERS4856645 SAMEA7096068 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096068 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#324 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#324 scientific_name Mus musculus strain C57BL/6J ERS4856646 SAMEA7096069 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096069 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#325 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#325 scientific_name Mus musculus strain C57BL/6J ERS4856647 SAMEA7096070 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096070 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#326 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#326 scientific_name Mus musculus strain C57BL/6J ERS4856648 SAMEA7096071 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096071 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#327 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#327 scientific_name Mus musculus strain C57BL/6J ERS4856649 SAMEA7096072 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096072 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#328 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#328 scientific_name Mus musculus strain C57BL/6J ERS4856650 SAMEA7096073 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096073 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#329 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#329 scientific_name Mus musculus strain C57BL/6J ERS4856651 SAMEA7096074 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096074 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#33 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#33 scientific_name Mus musculus strain C57BL/6J ERS4856652 SAMEA7096075 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096075 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#330 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#330 scientific_name Mus musculus strain C57BL/6J ERS4856653 SAMEA7096076 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096076 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#331 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#331 scientific_name Mus musculus strain C57BL/6J ERS4856654 SAMEA7096077 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096077 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#332 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#332 scientific_name Mus musculus strain C57BL/6J ERS4856655 SAMEA7096078 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096078 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#333 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#333 scientific_name Mus musculus strain C57BL/6J ERS4856656 SAMEA7096079 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096079 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#334 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#334 scientific_name Mus musculus strain C57BL/6J ERS4856657 SAMEA7096080 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096080 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#335 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#335 scientific_name Mus musculus strain C57BL/6J ERS4856658 SAMEA7096081 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096081 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#336 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#336 scientific_name Mus musculus strain C57BL/6J ERS4856659 SAMEA7096082 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096082 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#337 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#337 scientific_name Mus musculus strain C57BL/6J ERS4856660 SAMEA7096083 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096083 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#338 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#338 scientific_name Mus musculus strain C57BL/6J ERS4856661 SAMEA7096084 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096084 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#339 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#339 scientific_name Mus musculus strain C57BL/6J ERS4856662 SAMEA7096085 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096085 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#34 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#34 scientific_name Mus musculus strain C57BL/6J ERS4856663 SAMEA7096086 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096086 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#340 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#340 scientific_name Mus musculus strain C57BL/6J ERS4856664 SAMEA7096087 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096087 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#341 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#341 scientific_name Mus musculus strain C57BL/6J ERS4856665 SAMEA7096088 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096088 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#342 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#342 scientific_name Mus musculus strain C57BL/6J ERS4856666 SAMEA7096089 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096089 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#343 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#343 scientific_name Mus musculus strain C57BL/6J ERS4856667 SAMEA7096090 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096090 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#344 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#344 scientific_name Mus musculus strain C57BL/6J ERS4856668 SAMEA7096091 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096091 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#345 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#345 scientific_name Mus musculus strain C57BL/6J ERS4856669 SAMEA7096092 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096092 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#346 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#346 scientific_name Mus musculus strain C57BL/6J ERS4856670 SAMEA7096093 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096093 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#347 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#347 scientific_name Mus musculus strain C57BL/6J ERS4856671 SAMEA7096094 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096094 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#348 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#348 scientific_name Mus musculus strain C57BL/6J ERS4856672 SAMEA7096095 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096095 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#349 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#349 scientific_name Mus musculus strain C57BL/6J ERS4856673 SAMEA7096096 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096096 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#35 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#35 scientific_name Mus musculus strain C57BL/6J ERS4856674 SAMEA7096097 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096097 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#350 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#350 scientific_name Mus musculus strain C57BL/6J ERS4856675 SAMEA7096098 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096098 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#351 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#351 scientific_name Mus musculus strain C57BL/6J ERS4856676 SAMEA7096099 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096099 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#352 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#352 scientific_name Mus musculus strain C57BL/6J ERS4856677 SAMEA7096100 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096100 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#353 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#353 scientific_name Mus musculus strain C57BL/6J ERS4856678 SAMEA7096101 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096101 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#354 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#354 scientific_name Mus musculus strain C57BL/6J ERS4856679 SAMEA7096102 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096102 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#355 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#355 scientific_name Mus musculus strain C57BL/6J ERS4856680 SAMEA7096103 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096103 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#356 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#356 scientific_name Mus musculus strain C57BL/6J ERS4856681 SAMEA7096104 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096104 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#357 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#357 scientific_name Mus musculus strain C57BL/6J ERS4856682 SAMEA7096105 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096105 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#358 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#358 scientific_name Mus musculus strain C57BL/6J ERS4856683 SAMEA7096106 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096106 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#359 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#359 scientific_name Mus musculus strain C57BL/6J ERS4856684 SAMEA7096107 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096107 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#36 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#36 scientific_name Mus musculus strain C57BL/6J ERS4856685 SAMEA7096108 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:53Z External Id SAMEA7096108 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:53Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#360 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#360 scientific_name Mus musculus strain C57BL/6J ERS4856686 SAMEA7096109 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096109 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#361 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#361 scientific_name Mus musculus strain C57BL/6J ERS4856687 SAMEA7096110 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096110 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#362 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#362 scientific_name Mus musculus strain C57BL/6J ERS4856688 SAMEA7096111 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096111 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#363 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#363 scientific_name Mus musculus strain C57BL/6J ERS4856689 SAMEA7096112 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096112 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#364 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#364 scientific_name Mus musculus strain C57BL/6J ERS4856690 SAMEA7096113 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096113 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#365 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#365 scientific_name Mus musculus strain C57BL/6J ERS4856691 SAMEA7096114 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096114 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#366 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#366 scientific_name Mus musculus strain C57BL/6J ERS4856692 SAMEA7096115 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096115 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#367 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#367 scientific_name Mus musculus strain C57BL/6J ERS4856693 SAMEA7096116 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096116 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#368 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#368 scientific_name Mus musculus strain C57BL/6J ERS4856694 SAMEA7096117 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096117 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#369 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#369 scientific_name Mus musculus strain C57BL/6J ERS4856695 SAMEA7096118 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096118 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#37 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#37 scientific_name Mus musculus strain C57BL/6J ERS4856696 SAMEA7096119 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096119 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#370 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#370 scientific_name Mus musculus strain C57BL/6J ERS4856697 SAMEA7096120 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096120 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#371 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#371 scientific_name Mus musculus strain C57BL/6J ERS4856698 SAMEA7096121 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096121 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#372 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol RLT_SMT sample name E-MTAB-93451596018447:31617_2#372 scientific_name Mus musculus strain C57BL/6J ERS4856699 SAMEA7096122 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096122 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#373 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#373 scientific_name Mus musculus strain C57BL/6J ERS4856700 SAMEA7096123 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096123 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#374 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#374 scientific_name Mus musculus strain C57BL/6J ERS4856701 SAMEA7096124 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096124 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#375 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#375 scientific_name Mus musculus strain C57BL/6J ERS4856702 SAMEA7096125 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096125 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#376 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#376 scientific_name Mus musculus strain C57BL/6J ERS4856703 SAMEA7096126 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096126 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#377 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#377 scientific_name Mus musculus strain C57BL/6J ERS4856704 SAMEA7096127 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096127 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#378 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#378 scientific_name Mus musculus strain C57BL/6J ERS4856705 SAMEA7096128 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096128 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#379 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#379 scientific_name Mus musculus strain C57BL/6J ERS4856706 SAMEA7096129 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096129 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#38 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#38 scientific_name Mus musculus strain C57BL/6J ERS4856707 SAMEA7096130 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096130 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#380 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#380 scientific_name Mus musculus strain C57BL/6J ERS4856708 SAMEA7096131 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096131 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#381 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#381 scientific_name Mus musculus strain C57BL/6J ERS4856709 SAMEA7096132 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096132 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#382 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#382 scientific_name Mus musculus strain C57BL/6J ERS4856710 SAMEA7096133 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096133 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#383 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#383 scientific_name Mus musculus strain C57BL/6J ERS4856711 SAMEA7096134 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096134 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#384 age 8 to 10 broker name ArrayExpress buffer RLT cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol RLT_Maxima sample name E-MTAB-93451596018447:31617_2#384 scientific_name Mus musculus strain C57BL/6J ERS4856712 SAMEA7096135 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096135 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#39 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#39 scientific_name Mus musculus strain C57BL/6J ERS4856713 SAMEA7096136 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096136 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#4 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#4 scientific_name Mus musculus strain C57BL/6J ERS4856714 SAMEA7096137 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096137 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#40 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#40 scientific_name Mus musculus strain C57BL/6J ERS4856715 SAMEA7096138 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096138 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#41 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#41 scientific_name Mus musculus strain C57BL/6J ERS4856716 SAMEA7096139 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096139 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#42 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#42 scientific_name Mus musculus strain C57BL/6J ERS4856717 SAMEA7096140 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096140 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#43 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#43 scientific_name Mus musculus strain C57BL/6J ERS4856718 SAMEA7096141 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096141 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#44 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#44 scientific_name Mus musculus strain C57BL/6J ERS4856719 SAMEA7096142 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096142 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#45 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#45 scientific_name Mus musculus strain C57BL/6J ERS4856720 SAMEA7096143 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096143 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#46 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#46 scientific_name Mus musculus strain C57BL/6J ERS4856721 SAMEA7096144 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096144 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#47 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#47 scientific_name Mus musculus strain C57BL/6J ERS4856722 SAMEA7096145 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096145 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#48 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#48 scientific_name Mus musculus strain C57BL/6J ERS4856723 SAMEA7096146 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096146 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#49 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#49 scientific_name Mus musculus strain C57BL/6J ERS4856724 SAMEA7096147 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096147 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#5 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#5 scientific_name Mus musculus strain C57BL/6J ERS4856725 SAMEA7096148 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096148 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#50 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#50 scientific_name Mus musculus strain C57BL/6J ERS4856726 SAMEA7096149 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096149 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#51 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#51 scientific_name Mus musculus strain C57BL/6J ERS4856727 SAMEA7096150 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096150 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#52 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#52 scientific_name Mus musculus strain C57BL/6J ERS4856728 SAMEA7096151 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096151 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#53 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#53 scientific_name Mus musculus strain C57BL/6J ERS4856729 SAMEA7096152 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096152 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#54 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#54 scientific_name Mus musculus strain C57BL/6J ERS4856730 SAMEA7096153 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096153 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#55 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#55 scientific_name Mus musculus strain C57BL/6J ERS4856731 SAMEA7096154 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096154 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#56 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#56 scientific_name Mus musculus strain C57BL/6J ERS4856732 SAMEA7096155 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096155 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#57 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#57 scientific_name Mus musculus strain C57BL/6J ERS4856733 SAMEA7096156 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096156 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#58 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#58 scientific_name Mus musculus strain C57BL/6J ERS4856734 SAMEA7096157 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096157 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#59 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#59 scientific_name Mus musculus strain C57BL/6J ERS4856735 SAMEA7096158 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096158 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#6 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#6 scientific_name Mus musculus strain C57BL/6J ERS4856736 SAMEA7096159 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096159 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#60 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#60 scientific_name Mus musculus strain C57BL/6J ERS4856737 SAMEA7096160 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096160 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#61 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#61 scientific_name Mus musculus strain C57BL/6J ERS4856738 SAMEA7096161 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096161 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#62 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#62 scientific_name Mus musculus strain C57BL/6J ERS4856739 SAMEA7096162 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096162 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#63 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#63 scientific_name Mus musculus strain C57BL/6J ERS4856740 SAMEA7096163 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096163 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#64 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#64 scientific_name Mus musculus strain C57BL/6J ERS4856741 SAMEA7096164 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096164 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#65 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#65 scientific_name Mus musculus strain C57BL/6J ERS4856742 SAMEA7096165 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096165 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#66 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#66 scientific_name Mus musculus strain C57BL/6J ERS4856743 SAMEA7096166 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096166 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#67 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#67 scientific_name Mus musculus strain C57BL/6J ERS4856744 SAMEA7096167 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096167 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#68 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#68 scientific_name Mus musculus strain C57BL/6J ERS4856745 SAMEA7096169 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096169 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#69 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#69 scientific_name Mus musculus strain C57BL/6J ERS4856746 SAMEA7096170 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096170 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#7 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#7 scientific_name Mus musculus strain C57BL/6J ERS4856747 SAMEA7096171 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096171 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#70 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#70 scientific_name Mus musculus strain C57BL/6J ERS4856748 SAMEA7096172 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096172 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#71 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#71 scientific_name Mus musculus strain C57BL/6J ERS4856749 SAMEA7096173 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096173 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#72 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#72 scientific_name Mus musculus strain C57BL/6J ERS4856750 SAMEA7096174 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096174 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#73 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#73 scientific_name Mus musculus strain C57BL/6J ERS4856751 SAMEA7096175 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096175 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#74 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#74 scientific_name Mus musculus strain C57BL/6J ERS4856752 SAMEA7096176 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096176 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#75 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#75 scientific_name Mus musculus strain C57BL/6J ERS4856753 SAMEA7096177 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096177 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#76 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#76 scientific_name Mus musculus strain C57BL/6J ERS4856754 SAMEA7096178 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096178 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#77 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#77 scientific_name Mus musculus strain C57BL/6J ERS4856755 SAMEA7096179 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096179 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#78 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#78 scientific_name Mus musculus strain C57BL/6J ERS4856756 SAMEA7096180 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096180 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#79 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#79 scientific_name Mus musculus strain C57BL/6J ERS4856757 SAMEA7096181 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096181 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#8 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#8 scientific_name Mus musculus strain C57BL/6J ERS4856758 SAMEA7096182 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096182 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#80 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#80 scientific_name Mus musculus strain C57BL/6J ERS4856759 SAMEA7096183 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096183 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#81 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#81 scientific_name Mus musculus strain C57BL/6J ERS4856760 SAMEA7096184 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096184 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#82 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#82 scientific_name Mus musculus strain C57BL/6J ERS4856761 SAMEA7096185 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096185 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#83 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#83 scientific_name Mus musculus strain C57BL/6J ERS4856762 SAMEA7096186 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096186 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#84 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#84 scientific_name Mus musculus strain C57BL/6J ERS4856763 SAMEA7096187 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096187 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#85 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#85 scientific_name Mus musculus strain C57BL/6J ERS4856764 SAMEA7096188 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096188 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#86 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#86 scientific_name Mus musculus strain C57BL/6J ERS4856765 SAMEA7096189 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096189 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#87 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#87 scientific_name Mus musculus strain C57BL/6J ERS4856766 SAMEA7096190 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096190 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#88 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#88 scientific_name Mus musculus strain C57BL/6J ERS4856767 SAMEA7096191 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096191 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#89 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#89 scientific_name Mus musculus strain C57BL/6J ERS4856768 SAMEA7096192 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096192 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#9 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#9 scientific_name Mus musculus strain C57BL/6J ERS4856769 SAMEA7096193 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:54Z External Id SAMEA7096193 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:54Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#90 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#90 scientific_name Mus musculus strain C57BL/6J ERS4856770 SAMEA7096194 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:55Z External Id SAMEA7096194 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:55Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#91 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#91 scientific_name Mus musculus strain C57BL/6J ERS4856771 SAMEA7096195 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:55Z External Id SAMEA7096195 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:55Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#92 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#92 scientific_name Mus musculus strain C57BL/6J ERS4856772 SAMEA7096196 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:55Z External Id SAMEA7096196 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:55Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#93 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#93 scientific_name Mus musculus strain C57BL/6J ERS4856773 SAMEA7096197 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:55Z External Id SAMEA7096197 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:55Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#94 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#94 scientific_name Mus musculus strain C57BL/6J ERS4856774 SAMEA7096198 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:55Z External Id SAMEA7096198 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:55Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#95 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#95 scientific_name Mus musculus strain C57BL/6J ERS4856775 SAMEA7096199 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:55Z External Id SAMEA7096199 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:55Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#96 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme Maxima immunophenotype live cell organism part spleen protocol Triton_Maxima sample name E-MTAB-93451596018447:31617_2#96 scientific_name Mus musculus strain C57BL/6J ERS4856776 SAMEA7096200 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:55Z External Id SAMEA7096200 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:55Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#97 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#97 scientific_name Mus musculus strain C57BL/6J ERS4856777 SAMEA7096201 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:55Z External Id SAMEA7096201 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:55Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#98 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#98 scientific_name Mus musculus strain C57BL/6J ERS4856778 SAMEA7096202 10090 Mus musculus Protocols: Cells were collected from cultured mouse ESC cell line. FACS was used to enrich the cells. When FACS sorting, take care of plate calibration/priming prior to single-cell deposition. If many plates are deposited at the same time, repeat the calibration/priming at least every 8 plates. Defrost lysis buffer plates prior to cells sorting, centrifuge (1,000g for 1 min at 4oC) and keep chilled on ice. After FACS sorting, centrifuge immediately (1,000g for 1 min at 4 oC) the plate containing cells and keep them chilled on ice or at -80 oC until further processing. Transfer 1.2 μl of normalized samples by the Zephyr instrument to the working Nextera Plate. Keep plates chilled and continue with the Nextera protocol. Prepare the Nextera master mix. Once the temperature reaches 4 oC, remove from the thermal cycler and centrifuge (1,000g for 1 min at 4 oC) the plate. Wash Mantis LV chip with water by dispensing 50-100 μl of water into the waste plate. Dispense 1.2 μl NT buffer into each well to neutralize the reaction. Nextera Index preparation Timing 30min Prepare the Indexes set mix (A-B-C-D) in 96-well Index Plate according to the manufacturer's instructions. Defrost and spin down the Index 1(i7) and Index 2 (i5). Arrange the index primers in the Index Plate Fixture as follows: Index 1 (i7) adapters in columns 1-12 in ascending or descending order. Index 2 (i7) adapters in columns A-H in ascending or descending order. Using a multichannel pipette, add 10 μl of i7 adapters down each row. Replace the vial cap with the new cap. Using the multichannel pipette, add 10 μl of i5 adapters down each column. Replace the vial cap with the new cap. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Keep premade plates with index barcodes chilled until use or in the freezer at -20 oC for long term storage. Dispense 2.5 μl of Index Adapters on the Zephyr instrument to the Nextera plate from step 55. Seal the plate and centrifuge (1,000g for 1 min at 4 oC). Nextera Libraries amplification Timing 50min Wash Mantis LV chip by dispensing 50-100 μl of water into the waste plate. Dispense 3.7 μl of NPM buffer by the Mantis dispenser to each well of the Nextera plate. Seal the plate and centrifuge (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. Once the temperature reaches 4 oC, remove from the thermal cycler. Centrifuge the plate (1,000g for 1 min at 4 oC), vortex at 1800g for 1min and centrifuge (1,000g for 1 min at 4 oC) again. ENA-FIRST-PUBLIC 2020-12-31T11:11:49Z ENA-LAST-UPDATE 2020-07-29T11:28:55Z External Id SAMEA7096202 INSDC center name EMBL-EBI INSDC first public 2020-12-31T11:11:49Z INSDC last update 2020-07-29T11:28:55Z INSDC status public Submitter Id E-MTAB-93451596018447:31617_2#99 age 8 to 10 broker name ArrayExpress buffer Triton cell type splenocyte common name house mouse developmental stage adult disease normal enzyme SMT immunophenotype live cell organism part spleen protocol Triton_SMT sample name E-MTAB-93451596018447:31617_2#99 scientific_name Mus musculus strain C57BL/6J