ddrad sequencing of Brassica napus: leaf at flowering
Modified versions of the Peterson et al. (2012) and Clark et al. (2014) protocols were used to construct the ddRAD libraries. Digestion for the preparation of the ddRAD libraries was carried out using HpyCH4IV (5 U) and Hinfl (5 U) in NEB CutSmart buffer. The digested DNA was ligated respectively to the unique barcoded and common adapters using T4 DNA ligase (Thermo Invitrogen, Carlsbad, USA). the samples were not pooled after ligation but individually purified and double size selected to enable enrichment of fragments between 250 bp and 800 bp. A 10 L aliquot of the size selected DNA was used for PCR amplification.