Constructing Genomic Libraries using Super-GBS

we adopted modified GBS methods (SuperGBS, following Qi et al. [43 (link)]) to construct libraries. Briefly, extracted DNA from each individual was digested with both PstI-HF and MspI (New England Biolabs, NEB) for 2 h at 37 °C and then 2 h incubation at 75 °C. The barcoded adapters and common adapter were respectively ligated on the PstI cut site and the MspI cut site of all samples by T4 DNA ligase (NEB), ligation was running at 22 °C for 2 h. Fragments smaller than 300 bp were removed using recovery system of improved magnetic bead. The recovered fragment was amplified by PCR using high-fidelity enzymes, before libraries were sequenced, the concentration of PCR product was tested by Qubit2.0 and it should be greater than 5 ng/ul. Final libraries were sequenced using Illumina Hiseq Xten, PE150 Platform.