Rad-seq of callinectes sapidus: muscle tissue
Genetic data consisted of two reduced-representation genomic libraries of blue crabs from different localities along the US coast. The libraries' preparation involved a modified version of the double-digest, restriction-associated DNA sequencing (ddRAD) protocol of Peterson et al. (2012). Each sample containing 200 ng of DNA was digested with PstI and MluCI restriction enzymes. After heat inactivation, restriction site-compatible barcoded Illumina specific adaptors were ligated with the cut DNA with the PstI cuts receiving barcode P5 adapters and the MluCI cuts receiving half of the P7 adapter. After a second heat inactivation, samples were pooled and precipitated. These pooled DNA samples were size selected for 250-500 bp inserts using the Pipping Prep method (Sage Science). Next, samples were amplified using PCR with 20 cyles and included a biotinylated P5 primer and an indexed P7 reverse primer. Then, these PstI-MluCI fragments were enriched with Dynabeads M-280 streptavidin magnetic beads (Thermofisher).