GSM4594635: HA36_TKO_DNMT3A2; Mus musculus; Bisulfite-Seq
Genomic DNA extraction, followed by phenol/chloroform extraction and ethanol precipitation. RNaseA treatment was performed for 30 min at 37C Prior library preparation unmethylated Lambda phage DNA and SssI-methylated T7 DNA were spiked-in to genomic DNA samples to control bisulphite conversion rates. For WGBS, 20ug of DNA was sonicated to ca 500bp using a Bioruptor Pico. For RRBS, 20ug of DNA was digested for 18h with MspI and separated on 2% agarose gels. Digested DNA was excised at 150-250 and 250-350 bp. Sonicated and digested DNA samples for WGBS and RRBS, respectively, were end-repaired and A-tailed using the NEB-Ultra Illumina library preparation kit (E7370). DNA fragments were ligated to methylated adapters (NEB E7535) following the NEB Ultra protocol. After adapter removal using Ampure xp beads (Beckman Coulter), DNA was converted using the Qiagen Epitect bisulphite conversion kit following the FFT sample protocol. After conversion library PCR amplification was performed with 10 cycles following NEB Ultra kit protocol, and cleaned up using Ampure xp beads.
GEO Accession
GSM4594635