GSM8023352: 160080; Homo sapiens; Bisulfite-Seq
8ml of blood was drawn into the ACD tube.  After Ficol density gradient centrifugation, PBMCs were separated isolated  and cryopreserved in FCS/DMSO. Genomic DNA was extracted from PBMCs using phenol-chloroform method (Guha et al, 2018).  500ng genomic DNA was sheared and subject to end-repair, A-tailing and ligated with methylated adaptors.  Purified libraries were hybridized to biotinylated probes and subjected for bisulfite conversion (Zymo Cat# D5030).  Captured DNA was PCR amplified with KAPA HiFi HotStart Uracil+ (Cat# KK2801) into a final TBS-seq library.  Library quality was evaluated using TapeStation with the high-sensitivity D1000 tape (Agilent Cat# 5067- 5584).  A comprehensive TBS-seq protocol is demonstrated in (Morselli et al, 2021).