GSM9153255: 4cell_WGBS_KO.Kdm2a_KO.Kdm2b_KO_rep8; Mus musculus; Bisulfite-Seq
Genetically hybrid four-cell embryos were first treated with acidic Tyrode's solution containing 0.01% polyvinyl alcohol (PVA) to remove the zona pellucida. The embryos were then washed in M2 medium followed by PBS containing 0.01–0.05% PVA, and subsequently lysed in RLT Plus buffer. Ten four-cell embryos were pooled into a single tube containing the lysis buffer. DNA from the lysate was subjected to bisulfite conversion using the EZ DNA Methylation-Direct Kit and purified with the PureLink Micro Kit. Bisulfite-converted DNA underwent five rounds of strand synthesis, followed by exonuclease treatment and second-strand synthesis. The resulting DNA was then used for library preparation using NEB UDI indices and the KAPA HiFi HotStart PCR Kit. Final libraries were purified using AMPure XP beads.