GSM5747970: MHC region enriched GM12878 rep3; Homo sapiens; Bisulfite-Seq
Cultured cells were harvested and embed into low melting point agrose.After in-gel cleavege, the MHC moleculars were seperated and enriched by Pulse field gel electrophoresis . 20ng enriched MHC was fragmented to an average size of 250 bp using a Covaris M220 ultrasonicator. End repair and methylated adaptor ligation was performed with NEBNext Ultra End Repair/dA-Tailing Module, Ligation Module and NEBNext Multiplex Oligos for Illumina.Samples then underwent bisulfite conversion using an EZ DNA Methylation kit (Zymo Research, Irvine, CA, USA), with modified single stranded DNA fragments amplified using a Kapa HiFi U + HotStart ReadyMix (Kapa Biosystems, Wilmington, MA, USA) with primers (NEBNext Multiplex Oligos for Illumina). A final size selection was performed to enrich the library to a range between 300 and 500 bp. Constructed libraries were evaluated on an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA) to assess quality and sequenced on an Illumina HiSeq X Ten using the 150-bp paired-end mode.