trnL amplicon sequenced for Individual Pig TX05
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.
SUB3097761
TX05.trnL