GSM2601279: mESC_dCas9-Cat3a_noGuide_dox_48h_WGBS; Mus musculus; Bisulfite-Seq
genomic DNA was purified using Phenol/Chloroform extraction WGBS: 100-200 ng of genomic DNA was fragmented using a Covaris S2 for 6 min according to the following program: duty cycle 5%; intensity 10; cycle per burst 200. The sheared DNA was purified using the DNA Clean & Concentrator kit from Zymo Research. Bisulfite conversion of DNA was then conducted using the EZ DNA Methylation-Gold kit (Zymo Research), eluting in 15 µl low TE buffer. To minimize loss during storage, bisulfite-converted DNA was immediately processed for generating WGBS libraries using the Accel-NGS Methyl-Seq DNA library kit (Swift Biosciences). The libraries were sequenced for 100-bp paired-end reads on an Illumina HiSeq 2500 sequencer. RRBS: Genomic DNA was quantified using a Qubit® 2.0 Fluorometer, and quality-assessed on an Agilient 2200 TapeStation D1000 ScreenTape.  RRBS was performed on 20ng of each sample using the NuGen Ovation® RRBS Methyl-Seq System.  Barcoded samples were pooled in groups of 8 immediately prior to Bisulfite Conversion with the Qiagen EpiTect® Fast Bisulfite Conversion kit.  Library pools were purified with a 1X Agencourt RNA XP bead clean-up and sequenced on an Illumina HiSeq® 2500 Sequencing System. All protocols were carried out according to manufacturer's recommendations except where specified.
GEO Accession
GSM2601279