SRX18355214 2021-DUC10 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842719 2021-DUC10 2021-DUC10 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355215 2021-DUC12 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842721 2021-DUC12 2021-DUC12 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355216 2021-DUC45 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842720 2021-DUC45 2021-DUC45 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355217 2021-DUC46 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842722 2021-DUC46 2021-DUC46 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355218 2021-DUC47 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842723 2021-DUC47 2021-DUC47 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355219 2021-DUC48 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842724 2021-DUC48 2021-DUC48 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355220 2021-DUC49 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842725 2021-DUC49 2021-DUC49 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355221 2021-DUC50 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842726 2021-DUC50 2021-DUC50 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355222 2021-DUC51 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842727 2021-DUC51 2021-DUC51 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355223 2021-DUC55 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842728 2021-DUC55 2021-DUC55 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355224 2021-DUC56 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842729 2021-DUC56 2021-DUC56 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355225 2021-DUC59 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842730 2021-DUC59 2021-DUC59 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355226 2021-DUC2 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842731 2021-DUC2 2021-DUC2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355227 2021-DUC6 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842732 2021-DUC6 2021-DUC6 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355228 2021-DUC62 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842733 2021-DUC62 2021-DUC62 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355229 2021-DUC69 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842734 2021-DUC69 2021-DUC69 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355230 SB448 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842735 SB448 SB448 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355231 SB450 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842736 SB450 SB450 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355232 2021-DUC3 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842737 2021-DUC3 2021-DUC3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355233 SB453 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842738 SB453 SB453 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355234 SB454 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842739 SB454 SB454 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355235 SB457-1 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842740 SB457-1 SB457-1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355236 SB458 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842741 SB458 SB458 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355237 SB462 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842742 SB462 SB462 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355238 2021-DUC7 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842743 2021-DUC7 2021-DUC7 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355239 2021-DUC8 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842745 2021-DUC8 2021-DUC8 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355240 SB409 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842744 SB409 SB409 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355241 SB413 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842746 SB413 SB413 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355242 SB417 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842747 SB417 SB417 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355243 SB421 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842748 SB421 SB421 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355244 SB427 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842749 SB427 SB427 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355245 2021-DUC27 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842750 2021-DUC27 2021-DUC27 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355246 SB465 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842751 SB465 SB465 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355247 SB466 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842752 SB466 SB466 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355248 SB469 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842753 SB469 SB469 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355249 SB471-1 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842754 SB471-1 SB471-1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355250 2021-DUC31 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842755 2021-DUC31 2021-DUC31 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355251 2021-DUC33 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842756 2021-DUC33 2021-DUC33 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355252 2021-DUC37 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842757 2021-DUC37 2021-DUC37 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355253 2021-DUC41 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842758 2021-DUC41 2021-DUC41 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355254 SB428 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842759 SB428 SB428 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355255 SB430 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842761 SB430 SB430 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355256 SB434 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842760 SB434 SB434 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355257 SB437 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842762 SB437 SB437 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355258 SB438 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842763 SB438 SB438 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355259 SB443 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842764 SB443 SB443 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355260 SB444 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842765 SB444 SB444 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355261 SB446 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842766 SB446 SB446 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX18355262 2021-DUC42 SRP409306 bp0 The 16S rRNA v4 locusgene was amplified using the forward and reverse primers F515/R806. Each reaction consisted of 5.0 L of template DNA, 7.5 L of HotStart MyTaq Master Mix (Manufacturer, Headquarters), 1.0 L of 10 nM forward and reverse primers (combined), and 1.5 L of sterile water for a total of 15 L per reaction. PCR conditions consisted of 94 C for 3 min, 30 cycles of 94 C for 45 s, 50 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. We pooled amplicon triplicates and cleaned the products using the GeneJet PCR Clean-up Kit UltraClean PCR Clean-up kit. The second PCR was conducted to attach Illumina adaptors and dual index for the barcodes. The PCR consisted of 5.0 L of clean amplicons, 7.5 L 2X MyTaq Master Mix, 1.0 L of 10 nM forward and reverse barcode primers (Hernandez-Gomez et al., 2017), and 1.5 L of water for a total of 15 L reactions. PCR conditions consisted of 94 C for 3 min, 5 cycles of 94 C for 45 s, 65 C for 60 s, and 72 C for 90 s, followed by 72 C for 10 min. The PCR products were quantified using a Qubit Fluorometer, pooled in equimolar amounts, then cleaned one more time using the GeneJet PCR Clean-up Kit. The barcoded samples were shipped in dry ice overnight to thesent to Cornell University Institute for Biotechnology to be sequenced on a MiSeq machine (manufacturer, headquarters) using a 250 bp paired end protocol and sequencing kit. We used the 250 bp SRS15842767 2021-DUC42 2021-DUC42 AMPLICON METAGENOMIC PCR Illumina MiSeq