title: trnL amplicon sequenced for Individual Pig FL81
design: We collected fecal material as either fresh scat collected from transects within 24 hours of defecation following the methodology of (Kierepka et al. 2016; doi:10.1002/jwmg.21102), or from fecal material taken directly from the colon of culled individuals. Genomic DNA from fecal swabs was extracted using the MoBio PowerSoil-htp 96-well Isolation Kit (Carlsbad, CA).  A portion of the chloroplast trnL intron was PCR amplified using the g (5'- GGGCAATCCTGAGCCAA-3') and h (5'- CCATTGAGTCTCTGCACCTATC-3') primers for the trnL gene (Taberlet et al. 2007; doi:10.1093/nar/gkl938), but modified to include appropriate barcodes and adapter sequences for Illumina multiplexed sequencing. Unique per-sample 12-bp error-correcting barcodes were used, as described in Caporaso et al. (2012).  Each PCR reaction was mixed per the Promega PCR Master Mix specifications (Madison, WI), with 2 uL of gDNA template for a reaction volume of 25 uL.  The theromocycling program used an initial step at 94 °C for 2 minutes, a final extension at 72 °C for 2 minutes and the following steps cycled 35 times: 2 minutes at 94 °C, 1 minute at 55 °C, and 30 seconds at 72 °C.
identifiers.namespace: SUB3097761
identifiers.id: FL81.trnL