title: GSM3610157: WGBS: BS_TAB_diff63_ctr_d9_4; Homo sapiens; Bisulfite-Seq
library_construction_protocol: Briefly, genomic DNA was isolated using DNeasy Blood and Tissue kit (Qiagen) according to manufacturer's procedure. The bisulfite reaction was performed using EZ DNA Methylation Gold kit on 100 ng of gDNA according to manufacturer protocol. Genomic DNA was mixed with 0.5% of unmethylated lambda DNA (Promega) for assessment of efficiency of bisulfite reaction. For TAB-seq, the TAB reaction was performed according to published protocol and genomic DNA was sonicated for 30 cycles of 30 sec ON and 30 sec OFF on low power using a Bioruptor (Diagenode). Ox-BS was prepared by CEGX company using TrueMethyl kit workflow. For WGBS, random primed DNA synthesis and tagging was performed on converted gDNA. Finally, DNA was PCR amplified to generate a library for high-throughput sequencing. For TAB-seq, DNA was end-repaired and the ends where 3' adenylated in order to facilitate adapter ligation. Size selection was performed using Agencourt AMPure XP (Beckman Coulter) beads and, after adapter ligation and size selection, DNA was bisulfite treated using EpiTect Bisulfite kit (Qiagen) and PCR amplified using custom primers WGBS of wildtype LUHMES-derived neurons at day 9 was performed by whole genome bisulfite sequencing using an Epignome Methyl-seq kit (Epicentre, Illumina).
identifiers.namespace: GEO
identifiers.id: GSM3610157