GSM5342542: RRBS_Ocun_fibro_5aza0; Oryctolagus cuniculus; Bisulfite-Seq
Genomic DNA was extracted using the AllPrep DNA/RNA Mini Kit (Qiagen). 100 ng of genomic DNA were digested 5 hours by MspI (Thermo Scientific), end-repaired and A-tailed with Klenow fragment exo- (Thermo Scientific), and ligated to methylated adapters with T4 DNA ligase (Thermo Scientific) in Tango 1X buffer. Size selection was performed by gel excision from a 3% agarose 0.5X TBE gel to select fragments ranging from 150 to 400 bp. DNA was purified using the MinElute gel extraction kit (Qiagen) and bisulfite-converted twice with the EpiTect bisulfite kit (Qiagen), following the manufacturer's instructions. Final libraries were amplified using the Pfu Turbo Cx hotstart DNA polymerase (Agilent) with 12 to 14 PCR cycles. The libraries were purified using Agencourt AmpureXP beads (Beckman-Coulter).
GEO
GSM5342542