GSM2587321: NSC [MBD-seq]; Homo sapiens; MBD-Seq
1ug of gDNA was sheared to 200~400bp using the Covaris LE220 sonicator (Covaris Inc. Woburn, MA). The fragmented products were immunoprecipitated using the MethylMiner Methylated DNA Enrichment Kit (Invitrogen) following the manufacture’s recommended protocol. Briefly, Methylated DNA was isolated from fragmented whole genomic DNA by binding to the methyl-CpG binding domain of human MBD2 protein, which is coupled to paramagnetic Dynabeads® M-280 Streptavidin via a biotin linker. The methylated fragments were eluted as a single enriched population with a 2000 mM NaCl elution buffer. DNA libraries were prepared for sequencing using MethylMiner Methylated DNA Enrichment Kit (Invitrogen).