GSM4082561: 101_E114F_father_CTRL; Homo sapiens; Bisulfite-Seq
Genomic DNA (gDNA) was extracted from stored whole blood samples using the QIAmp DNA Blood Mini Kit (Qiagen, cat.: 51106), according to manufacturer's instruction and stored in -80 ℃ if not used immediately. In brief, approximately 0.3% unmethylated lambda DNA (dam-; dcm-) (Thermo Scientific, cat.: SD0021) was spiked into 500 ng of high-quality gDNA, and the mixture was digested by MspI (Thermo Scientific, cat.: ER0541). After purification with Agencourt AMPure XP beads (Agencourt, cat.: A63881), the digested DNA was end-repaired and tailed with deoxyadenosine using Klenow Fragment exo- (Thermo Scientific, cat.: EP0422). Then, NEBNext adapters (NEB, cat.: E7335) were ligated overnight with T4 DNA ligase at a high concentration (NEB, cat.: M0202M) with subsequent digestion by Uracil-Specific Excision Reagent (USER) enzyme (NEB, cat.: M5505L).