title: GSM8163650: oocytes, SN_rep2; Mus musculus; Bisulfite-Seq
library_construction_protocol: Collect 20 oocytes into 2.5 μL of Buffer RLT Plus (Qiagen, 1053393) for cell lysis. The construction of BS-seq libraries was adapted from previously outlined method(47), with some adjustments.Incubate the lysate with 65 μL of CT conversion reagent (Zymo, d5455) for 8 minutes at 98°C, followed by 180 minutes at 65°C. The BS-converted DNA, purified with Agencourt AMPure XP beads, received preamp oligo and 50 U of Klenow exo- polymerase addition for preamplification, which was then subjected to exonuclease I treatment and subsequent preamplification purification. Double-tagged products were generated through PCR with adapter oligo. Finally, the libraries were constructed by a 15-cycle of PCR amplification using Illumina universal indexed primers.