RAD-seq of kiwifrui
Briefly, genomic DNA from the female parent and each of 102 progenies were digested for 15 min at 37буC in a 50 ж╠L reaction with 20 units (U) of EcoR I (NEB, USA). P1 adapter, a modified Illumina adapter, was ligated to the samples. After adapter ligation, sample were pooled and randomly sheared with a Bioruptor (Diagenode, Belgium) into an average size of 500 bp. DNA fragments in the range of 300иC500 bp were purified using the MinElute Gel Extraction Kit (Qiagen). The dsDNA ends were repaired using the Quick Blunting kit Enzyme Mix (NEB). Then, a modified Solexa P2 adapter was ligated to the obtained DNA fragments. Finally, purified and quantified DNA products were PCR-amplified using Phusion Master Mix (NEB, USA). The prepared DNA libraries were sequenced on the Illumina Hiseq2000 instrumentation.
F13FTSCCKF2054