ERS2647415
SAMEA4827534
Sample 1
10090
Mus musculus
Protocols: Stable GM40600- and EGFP-expressing SP 2/0 cells, and the control EGFP-expressing SP 2/0 cells were collected after after cultured for 2 days in fresh medium. Samples were washed twice with PBS and 1 mL of TRIzol per 1 x 107 cell pellets was added and frozen in -80℃. GM40600 cDNA (General Biosystems, Anhui, China) was cloned into lentiviral vector EGFP-expressing LV201 (Fugene Corp., Guangzhou, China) to generate GM40600 protein. Empty LV201 and GM40600-expressing LV201 was transfected into SP 2/0 cells (ATCC® CRL-1581™, https://www.atcc.org/products/all/CRL-1581.aspx), and stable transfectants were identified by EGFP expression and drug selection (Puromycin, Sigma, 10 μg/ml). Stable GM40600-expressing SP 2/0 cells and LV201-transduced SP 2/0 cells were cultured in fresh ATCC-formulated Dulbecco's Modified Eagle's Medium (Catalog No. 30-2002) containing 10% FBS, 2 mM glutamine, penicillin (100 IU/ml), streptomycin (100 ug/ml), and 50 mM 2-mercaptoethanol. Cells were collected on ice after cultured for 2 days in fresh medium. Data were aligned using STAR with options --outFilterScoreMinOverLread 0.4 --outFilterMatchNminOverLread 0.4, against the C3H/HeJ genome as a reference (ftp://ftp.ensembl.org/pub/release-89/fasta/mus_musculus_c3hhej/dna/Mus_musculus_c3hhej.C3H_HeJ_v1.dna_sm.toplevel.fa.gz). Total RNA from cells was isolated and purified using the RNeasy Mini Kit (Qiagen, Venlo, Netherlands). All RNA content in the samples was measured with a NanoDrop®ND-1000 spectrophotometer, and the RNA quality was assessed using RNA 6000 NanoChips with the Agilent 2100 Bioanalyzer (Agilent, Palo Alto, CA, USA). Libraries were prepared using the TruSeq Stranded Total RNA Library Prep Kit with Ribo-Zero Gold (Illumina).
ENA-FIRST-PUBLIC
2019-07-15T04:04:45Z
ENA-LAST-UPDATE
2018-08-13T11:13:02Z
External Id
SAMEA4827534
INSDC center name
Beijing Institute of Basic Medical Sciences
INSDC first public
2019-07-15T04:04:45Z
INSDC last update
2018-08-13T11:13:02Z
INSDC status
public
Submitter Id
E-MTAB-7136:Sample 1
broker name
ArrayExpress
cell line
Sp2/0-Ag14
cell type
lymphocyte
common name
house mouse
disease
multiple myeloma
genotype
empty vector LV201
organism part
mixed spleen and skin
phenotype
empty vector
sample name
E-MTAB-7136:Sample 1
scientific_name
Mus musculus
ERS2647416
SAMEA4827535
Sample 2
10090
Mus musculus
Protocols: Stable GM40600- and EGFP-expressing SP 2/0 cells, and the control EGFP-expressing SP 2/0 cells were collected after after cultured for 2 days in fresh medium. Samples were washed twice with PBS and 1 mL of TRIzol per 1 x 107 cell pellets was added and frozen in -80℃. GM40600 cDNA (General Biosystems, Anhui, China) was cloned into lentiviral vector EGFP-expressing LV201 (Fugene Corp., Guangzhou, China) to generate GM40600 protein. Empty LV201 and GM40600-expressing LV201 was transfected into SP 2/0 cells (ATCC® CRL-1581™, https://www.atcc.org/products/all/CRL-1581.aspx), and stable transfectants were identified by EGFP expression and drug selection (Puromycin, Sigma, 10 μg/ml). Stable GM40600-expressing SP 2/0 cells and LV201-transduced SP 2/0 cells were cultured in fresh ATCC-formulated Dulbecco's Modified Eagle's Medium (Catalog No. 30-2002) containing 10% FBS, 2 mM glutamine, penicillin (100 IU/ml), streptomycin (100 ug/ml), and 50 mM 2-mercaptoethanol. Cells were collected on ice after cultured for 2 days in fresh medium. Data were aligned using STAR with options --outFilterScoreMinOverLread 0.4 --outFilterMatchNminOverLread 0.4, against the C3H/HeJ genome as a reference (ftp://ftp.ensembl.org/pub/release-89/fasta/mus_musculus_c3hhej/dna/Mus_musculus_c3hhej.C3H_HeJ_v1.dna_sm.toplevel.fa.gz). Total RNA from cells was isolated and purified using the RNeasy Mini Kit (Qiagen, Venlo, Netherlands). All RNA content in the samples was measured with a NanoDrop®ND-1000 spectrophotometer, and the RNA quality was assessed using RNA 6000 NanoChips with the Agilent 2100 Bioanalyzer (Agilent, Palo Alto, CA, USA). Libraries were prepared using the TruSeq Stranded Total RNA Library Prep Kit with Ribo-Zero Gold (Illumina).
ENA-FIRST-PUBLIC
2019-07-15T04:04:45Z
ENA-LAST-UPDATE
2018-08-13T11:13:02Z
External Id
SAMEA4827535
INSDC center name
Beijing Institute of Basic Medical Sciences
INSDC first public
2019-07-15T04:04:45Z
INSDC last update
2018-08-13T11:13:02Z
INSDC status
public
Submitter Id
E-MTAB-7136:Sample 2
broker name
ArrayExpress
cell line
Sp2/0-Ag14
cell type
lymphocyte
common name
house mouse
disease
multiple myeloma
genotype
LV201_Gm40600
organism part
mixed spleen and skin
phenotype
Gm40600 overexpression
sample name
E-MTAB-7136:Sample 2
scientific_name
Mus musculus