Genotyping-by-sequencing of North American squamates
Genotyping-by-sequencing (GBS) libraries were prepared following a modified version of the protocol described in Elshire et al. (2011). We digested 100 ng of input DNA with the enzyme Pst1, ligated 8 L of adapter mix including unique barcodes, pooled libraries, and performed a bead cleanup with Sera-Mag Speedbeads (Rohland and Reich 2012). Final PCR amplification was conducted with 16 cycles in 8 replicate reactions, followed by pooling, bead cleanup, and quantification via a Qubit fluorometer (Life Technologies). Libraries were size selected to 200-500 bp using a Blue Pippen (Sage Science Inc.), quantified with a Bioanalyzer (Agilent Technologies), and sequenced at the OSU Comprehensive Cancer Center Genomics Shared Resource on an Illumina HiSeq 4000 with 150 bp sequencing