population RAD-seq for Agriophyllum squarrosum
1g genomic DNA was digested for 3-4 hours at 37 in a 40 L reaction with 20 units (U) of EcoRI restriction enzyme (specific for the G|AATTC motif) per sample. Then, the digested and purified DNA was ligated to P1 barcoded adapters, and was sheared to an average size of 350 bp using a Biorupter pico. After P2 adapter ligation, to remove adapter dimers, libraries with DNA 300-500 bp were further purified using Qiagen Mini Elute Kit (Qiagen) according to the protocol of the manufacturer. All the libraries were enriched with PCR amplification using KAPA Library Amplification Kits, and then purified again with Qiagen Mini Elute Kit, and then sequenced on a HiSeq 3000 platform (Illumina, San Diego, CA, USA) as 150 bp paired end sequences with one library per lane.