title: 16S sequencing of feces and foregut contents of captive and wild Amargosa voles
design: 16S rRNA gene V3-V4 variable region PCR primers 515/806 with barcode on the forward primer were used in a 30 cycle PCR using the HotStarTaq Plus Master Mix Kit (Qiagen) under the following conditions: 94oC for 3 minutes, followed by 28 cycles of 94C for 30 seconds, 53C for 40 seconds and 72C for 1 minute, after which a final elongation step at 72C for 5 minutes was performed. After amplification, PCR products were examined by agarose gel electrophoresis for success of amplification and relative band intensity. Multiple samples were pooled together in equal proportions (based on their molecular weight and DNA concentrations), purified using calibrated Ampure XP beads, and then used to prepare a DNA library using the Illumina TruSeq DNA library preparation protocol