GSM5351783: 256_cell_dnmt1_mKD_BS_seq; Danio rerio; Bisulfite-Seq
The deyolked oocytes were lysed with 20 µl lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1 mM EDTA, pH 8.0, 0.5% NP-40) and 2 µl protease K (Roche, Cat 10910000) for at least 3hrs at 55℃. After heat-inactivation, spike-in λ-DNA (Promega, Cat D150A) was added at a mass ratio of 1/200. Bisulfite conversion was performed with the EpiTect Fast Bisulfite Conversion Kit (Qiagen, Cat 59824). The converted DNA was subjected to column purification and desulfonation on MinElute DNA spin columns (Qiagen, Cat 59824) with carrier RNA (Qiagen, Cat 59824) according to the manufacturer's instructions. The purified DNA was eluted in 30 µl of elution buffer and ready for TELP library preparation (Peng et al., 2015).