GSM8080151: RH RRBS VEH-1 replicate; Mus musculus; Bisulfite-Seq
The genomic DNA was extracted from mouse HSPCs per condition using QIAamp DNA extraction kit. Genomic DNA was digested with MspI, and the resulting fragments were end-repaired, A-tailed, and ligated with methylated adapters. After size-selection, bisulfite conversion was performed using the EZ DNA Methylation-Gold™ Kit (Zymo Research). The bisulfite-converted DNA was PCR-amplified to enrich for adapter-ligated fragments. Size selection was conducted via gel cutting in order to obtain DNA fragments spanning a range of 220bp to 40bp in length (Meissner, 2008). Following this, the EZ DNA Methylation Gold Kit (Zymo Research) was utilized to convert these fragments to bisulfite. The ultimate DNA library was generated via PCR amplification.