title: GSM5982013: RRBS_Jurkat-SV_DMSO_rep2; Homo sapiens; Bisulfite-Seq
library_construction_protocol: For RRBS, 1 x 106 cells were lysed, incubated with RNAse A and Proteinase K, and genomic DNA was purified using Monarch Genomic DNA purification kit (New England Biolabs). 0.5 - 1 ug of genomic DNA was digested with MspI for 2 hours and digested DNA was extracted using phenol/chloroform and purified by ethanol precipitation. MspI-digested DNA was incubated with Klenow Fragment, exo- (New England Biolabs) and dNTPs (10 mM dATP, 1 mM dGTP, 1 mM dCTP) for 30 min at 30°C  followed by 30 min at 37°C to perform end-repair and A-tailing, respectively. Methylated Illumina adapter (New England Biolabs) was ligated to end-repaired, A-tailed using T4 DNA ligase followed by digestion with USER enzyme mix (New England Biolabs). Size selection of 40-220bp inserts was performed by gel extraction from 3% NuSieve 3:1 (Lonza) agarose gel. Extracted DNA libraries underwent two consecutive rounds of bisulfite conversion using EpiTect Bisulfite kit (QIAGEN). Final RRBS libraries were constructed by PCR using NEBNext Index Primers for Illumina (New England Biolabs) and PfuTurbo Cx Hotstart polymerase (Agilent Technologies).