ddRADSeq of Peristenus
the enzyme pair NlaIII and MluCl (NEB, Ipswich, MA, USA) were chosen. Libraries were prepared on 48 individuals grouped by DNA yield, with each sample assigned one of 48 unique 5-base pair (bp) in-line barcode sequences during adapter ligation. Each set of 48 samples was then pooled for automated selection of a 216-336bp fragments on a PippinHT (Sage Science, Beverly, MA, USA), followed by 12 rounds of high-fidelity PCR amplification (Q5 High-Fidelity DNA Polymerase, NEB) using PCR primers that included one of 12 unique Illumina multiplex read indices.