title: GSM3213920: OxBS_DKO_72h; Mus musculus; Bisulfite-Seq
library_construction_protocol: DNA was purified using DNaeasy Qiagen blood and tissue kit (Qiagen). Three PCR products containing C, mC, or hmC pertaining to different regions of ʎ phage genome were used as spike-ins at a ratio of 1:200 of the genomic DNA. 1.5 µg of genomic DNA mixed with spike-ins was precipitated with ethanol and 1 µg of the precipitated DNA was oxidized with potassium perruthenate (KRuO4; Sigma) followed by bisulfite treatment (for oxBS). As control, 0.5 µg of DNA was directly used for bisulfite treatment. The BS and oxBS treated DNA were amplified using respective PCR primers and as well as primers specific to the spike-in PCR products with KAPA Uracil+ PCR mix (Roche). The amplified products were pooled and libraries were prepared using the NEB Ultra II library preparation kit (NEB) according the manufacturer. Bisulfite-seq, paired-end 250 with Miseq
attributes.tag: GEO Accession
attributes.value: GSM3213920