alias: GSM3303957
title: GSM3303957: XWGBS_GM12884; Homo sapiens; Bisulfite-Seq
library_construction_protocol: 2 million cells were pelleted at 2000g for 10' and frozen at -80C until extraction of DNA. Pellets were thawed and resspended in 10mM Tris pH 8.0, 150 mM EDTA solution before the addition of SDS to a final concentration of 1%. Protein was digested overnight at 50C with 25 ug of Proteinase K, followed by an hour incubation at 37C with 5 ug of RNAse A. Extraction of genomic DNA was then achieved through two consecutive phenol:chloroform steps, selecting the aqeous phase. The DNA was then concentrated and purified via ethanol precipitation. 100 ng of genomic DNA was treated with Tn5 transposase for tagmentation, then tagged DNA was filled with methylated dCTP, dATP, dGTP, dTTP mixture. The filled DNA was bisulphite treated using EZ DNA methylation Gold kit (Zymo research). The bisulphite treated samples were amplified with custom and Illumina index adapters. The amplified products were size selected using AMPureXP beads.
identifiers.namespace: GEO
identifiers.id: GSM3303957