ddRADseq leopard seal
Genomic DNA was digested with the restriction endonucleases PstI and MseI and processed into SBG libraries. Briefly, genomic DNA was digested in a reaction with PstI and MseI (New England Biolabs). After digestion, the samples were heat-inactivated and P1 and P2 adaptors were added. Next, the samples were diluted and part of this product was used in a PCR amplification utilizing MseI and PstI primers. The PCR product obtained from each sample was pooled, purified and eluted and ran on a 1.5% agarose gel. DNA bands appearing at 300 bp to 800 bp were excised from the gel and purified with a gel extraction kit. The excised DNA was again subjected to restriction digestion with SgrAI. Then, individual barcodes and sequence adapters were ligated to the genomic DNA. The barcoded samples were pooled, and fragments were sequenced to obtain 100bp single end reads in a HiSeq 2000 platform (Illumina). The reads were de-multiplexed and barcodes were trimmed resulting in final DNA fragment lengths of 95 bp.