title: 16S rRNA gene seq of homo sapiens: adult stool: Sample S10531
design: DNA was extracted from stool using the PowerLyzer PowerSoil Kit (Mo Bio Laboratory Inc., CA) following manufacturer’s protocol. Barcoded amplicons were generated covering the V4 region of the 16S rRNA gene using the F515/R806 primer pair. The PCR reaction, using FastStart High Fidelity PCR system, dNTP pack (Roche, IN), was run as follows: initial denaturing at 94C for 3min, followed by 25 cycles of 94C for 15s, 52C for 45s and 72C for 1min, and a final extension at 72C for 8min. PCR products were purified using Agencourt AMPure XP (Beckman Coulter Life Sciences, IN) and quantified using the Agilent 4200 TapeStation (Agilent Technologies, CA). Amplicon libraries were pooled at equal molar concentrations and sequenced on Illumina MiSeq with a 300-cycle (2x151 bp) kit.