DNA was isolated from the 2-month-old castor bean plants of each of the 200 RILs. DNA extraction was carried out using the plant genomic DNA extraction Kit (TIANGEN, Beijing, China) following the manufacturer’s instructions. RNase A was then added to digest RNA. The quality and concentrations of DNA were detected using a NanoDrop 2000 (Thermo Fisher Scientific, USA), while DNA integrity was examined by electrophoresis on 1% agarose gels.
To prepare the reduced representation libraries for sequencing, the GBS protocol was carried out according to the method reported by Elshire et al.17 (link). In brief, the genomic DNA was first digested by restriction enzymes. In this case, EcoRI and MseI were selected to efficiently reduce genome complexity. The barcode adapters and common adapters were then linked to the sequence ends of the fragmented DNA samples. PCR amplification and fragment selection was performed. Libraries were then sequenced by Illumina HiSeq3000 platform, which generated 150 bp paired-end reads.
To prepare the reduced representation libraries for sequencing, the GBS protocol was carried out according to the method reported by Elshire et al.17 (link). In brief, the genomic DNA was first digested by restriction enzymes. In this case, EcoRI and MseI were selected to efficiently reduce genome complexity. The barcode adapters and common adapters were then linked to the sequence ends of the fragmented DNA samples. PCR amplification and fragment selection was performed. Libraries were then sequenced by Illumina HiSeq3000 platform, which generated 150 bp paired-end reads.
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