GSM5411616: hSMC_5hmC_IP; Homo sapiens; OTHER
hydroxymethylated DNA was immunoprecipitated by Hydroxymethylated DNA IP Kit (Diagenode) following manufacturer's protocol. Precipitated genomic DNA was sonicated to 200 – 500 bp using the Covaris sonicator. 1 µg of fragmented genomic DNA was immunoprecipitated with a 5-hmC monoclonal antibody using the hMeDIP kit (Diagenode). The DNA-antibody mixture was then incubated with magnetic beads overnight at 4°C, washed and DNA purified using purification kit (Qiagen, 69504) DNA library was prepared by NEBNext DNA library prep Master Mix set for illumina 6040 and NEBNext Multiplex Oligos for Illumina (Index Primers Set 1) E7335.