whole genome Bisulfite-Seq ofYak-Lung-Rep2
1 g of genomic DNA was fragmented by sonication to a mean size of approximately 250 bp, and subsequently used for whole genome bisulfite sequencing (WGBS) library construction using Acegen Bisulfite-Seq Library Prep Kit (Acegen) following the manufacturers instructions. Briefly, fragmented DNA was end-repaired, 5'-phosphorylated, 3'-dA-tailed and then ligated to methylated adapters. The methylated adapter-ligated DNAs were purified using 1 Agencourt AMPure XP magnetic beads and subjected to bisulfite conversion by ZYMO EZ DNA Methylation-Gold Kit (zymo). The converted DNAs were then amplified using 25 l HiFi HotStart U+ RM and 8-bp index primers with a final concentration of 1 M each. The constructed WGBS libraries were then analyzed by Agilent 2100 Bioanalyzer and quantified by a Qubit fluorometer with Quant-iT dsDNA HS Assay Kit (Invitrogen), and finally sequenced on Illumina Hiseq X ten sequencer.