GSM1545748: TKO_DNMT3B1_SETD2_bisulfite_seq single; Mus musculus; Bisulfite-Seq
Genomic DNA was extracted using Proteinase K digest followed by phenol extraction and ethanol precipitation, followed by RNase treatment. Genomic DNA was sonicated using a Bioruptor (7 cycles) to 200-500 bp. Fragmented, pre-methylated phage T7 DNA and unmethylated lambda DNA were spiked in as converson control. 6 ug DNA were end-repaired, poly-adenylated and ligated to methylated, paired-end adapters from Illumina. Ligated DNA fragments were purified on 2% agarose gels. Fragmetns between 350-400bp were extracted and bisulfite conversion was performed using the Qiagen Epitect bisulfite kit. Converted DNA was amplified for 10 cycles and cleaned up with Ampure beads.