GSM8151240: 160013; Homo sapiens; Bisulfite-Seq
500 ng of extracted DNA were used for TBS-seq library preparation. Fragmented DNA was subject to end repair, dA-tailing and adapter ligation using the NEBNext Ultra II Library prep kit using custom pre-methylated adapters (IDT). Pools of 16 purified libraries were hybridized to the biotinylated probes according to the manufacturer's protocol. Captured DNA was treated with bisulfite prior to PCR amplification using KAPA HiFi Uracil+(Roche) with the following conditions: 2 min at 98◦C; 14 cycles of (98◦C for 20 sec; 60◦C for 30 sec; 72◦C for 30 sec); 72◦C for 5 minutes; hold at 4◦C. Library QC was performed using the High-Sensitivity D1000 Assay on a 2200 Agilent TapeStation. Pools of 96 libraries were sequenced on a NovaSeq6000 (S1 lane) as paired-end 150 bases.