title: GSM7729297: SSSavi_DKE_WGBS_C; Homo sapiens; Bisulfite-Seq
library_construction_protocol: WGBS: gDNA was extracted using a Qiagen DNeasy Blood & Tissue Kit (#69504). From this, 200 ng of each sample and 2 ng of lambda DNA (Promega #D150A) was sheared (Covaris, M220) for 5 min, 20% duty cycle, 75 peak power, and 200 cycles per burst. Sheared DNA fragments were then End-repaired and A-tailed using a NxSeq AmpFREE Low DNA Lib Kit (Lucigen, #14000-2), followed by ligation of Nextflex methylated adapters (Perkin Elmer, #NOVA-511913) and cleaned up and size selected for >300 bp using Agencourt AMPure XP beads. Samples were then bisulfite converted using a EZ DNA Methylation-Direct Kit (Zymogen, #D5021) following the manufacturer's protocol.  KAPA HiFi Uracil+ readymix (Roche, #KM2801) was then used for 6 cycles of library amplification using 25 µM mix of p5 and p7 primers (5' AATGATACGGCGACCACCGAGATCTACAC and 5' CAAGCAGAAGACGGCATACGAGAT, respectively) primrs from IDT, followed by a final bead clean-up.