GSM2254279: Sall4 ko EAF oocyte, replicate 1 [RRBS-seq]; Mus musculus; Bisulfite-Seq
Single oocyte harvested from Sall4fl/fl;Zp3-Cre and Sall4fl/fl female mice at P12.5 and P17.5 The DNA from the nuclei of the oocytes were extracted, with digestion of nuclear proteins. The DNA was further treated with MspI digestion (Fermentas), end-repair, dA-tailing, adaptor-ligation, and bisulfite-conversion in a one-tube reaction. The MethylCode Bisulfite Conversion Kit (Invitrogen, MECOV-50) was used for bisulfite-conversion and purification. Unmethylated lambda DNA (Fermentas) was added to the gDNA sample before MspI digestion to monitor the bisulfite conversion rate. The converted libraries were further purified with Agencourt AMPure XP beads (Beckman A63881) and amplified by 2-round PCR enrichment. After amplification, size-selection was performed on the 12% native polyacrylamide TBE gel to obtain the 200–500 bp DNA fragments.