GSM7508588: oocyte from antral follicle stage 1, gilt 201, rep3 [BS-Seq]; Sus scrofa; Bisulfite-Seq
Firstly, single oocyte in cell lysis buffer was vortexed for 2 min then centrifugated at 1200 g for 10 min at 4 ℃, resulting in the physical separation of mRNA and genomic DNA. The sediment containing nuclei was subjected to single-cell bisulfite scWGBS library construction. Bisulfite conversion was carried out using EZ Methylation Direct bisulfite reagent (Zymo) on purified DNA in the presence of AMPure XP beads. BS-converted DNA was then submitted to preamplification using primers contained a 3'-random nonamers, and adaptors were incorporated during a total of five times of random priming and extension.  An extra 0.8 x AMPure XP purification was performed before final PCR amplification, then the purified libraries were ready for quality control and sequencing.