Rattus rattus fecal DNA metabarcoding: 16S amplicons
DNA was extracted from 1g rat stool using Zymo Quick-DNA Fecal/Soil Microbe Miniprep kits._x000D_   16S metabarcoding was conducted using 515F806R primers to target the V4 region of the 16S SSU rRNA. _x000D_   Each primer included an Illumina adapter, barcode, primer pad, and linker. Reactions were carried out in 25uL volumes_x000D_   consisting of 10uL of 1.25uM forward and reverse primer, 2uL of DNA, and 13uL of Platinum Hot Start PCR mastermix _x000D_   (ThermoFisher Scientific, cat 13000014). Reaction conditions were as follows: 95C for 3min, 35x 98C for 30secs, _x000D_   58C for 30secs, and 72C for 30secs, followed by a final extension at 72C for 5min. Concentrations were measured using _x000D_   Promega One Quantifluor kits on a Teccan platereader. Samples were then normalized to 7ng/uL prior to pooling. _x000D_   The product was cleaned using magnetic beads (bead:DNA ratio was 0.8:1) and sequenced by sampling village at UC Santa Barbara Biological _x000D_   Nanostructures Laboratory on an Illumina MiSeq (v3 chemistry, 2x300bp, 24M reads).