ddRAD-seq of Spea multiplicata: juvenile toe clip
At metamorphosis, a toe clip was taken and stored in ethanol at -80C. Genomic DNA was extracted using a Qiagen DNeasy Blood & Tissue Kit (Qiagen Inc.). Double digest RAD-seq libraries were built using the enzyme pair SphI and MluCI following previously published protocols. Samples were distributed across four libraries using 200 ng of template DNA per individual. Single-end 100 bp sequencing was performed on two lanes (two libraries per lane) of an Illumina HiSeq 4000 at the University of Oregon GC3F facility.