title: RAD-seq of Pungitius laevis: female
design: Genomic DNA was extracted from ethanol preserved fin clips or muscle tissue using the standard phenol-chloroform method. Approximately 100 ng of genomic DNA was isolated for RAD library construction following the protocol of Baird et al. (2008). DNA was fragmented with restriction enzyme (PstI). P1 adapter with forward amplification and sequencing primers and sample specific barcode were ligated to the digested DNA fragments. Barcoded samples were sheared randomly and the P2 adapter was added. DNA fragments of 300 to 500 bp in length were gel purified after selective PCR enrichment step. Barcoded RAD samples were then pooled and sequenced using the Illumina HiSeq2000 platform with 100 bp paired-end strategy.