title: ddRADseq of Pelophylax esculentus complex families: parents and progeny
design: template DNA for each sample at equimolar concentrations (approx. 15 ng/ul) was digested using a combination of MspI and SbfI restriction enzymes. P1 and P2 adapters containing barcodes were then ligated, and these ligated fragments were amplified in four PCR replicates of 20 cycles, lowering the risk of polymerase-induced errors. The products were then pooled, and fragments of 350 - 500 bp were selected with 2% agarose gel, of which 135-139 bp belonged to the pair of Illumina primers, adaptors, and barcodes. The pool of 96 individual libraries was sequenced with a single-end run of 100 cycles (for 150 bp) on a HiSeq 2500 Illumina instrument at the Genomic Technologies Facility at the University of Lausanne, Switzerland.