GSM5417973: Hi-C AB1 WT rep2; Mus musculus; Hi-C
Crosslinked cells were lysed in ice-cold Hi-C lysis buffer and treated with SDS. SDS was then quenched with Triton X-100. Chromatin was digested with MboI restriction enzyme and digested overhang was labeled with biotin-14-dATP. DNA fragments were ligated. After that, crosslinking was reversed and DNA was purified. DNA was sheared to 300-500 bp and biotin-labeled DNA was enriched using Dynabeads MyOne Streptavidin T1 beads. The following library construction steps were performed on beads. Briefly, sheared DNA ends were repaired and ligated with dATP tailing. Then Illumina adapters were ligated to DNA fragments and 8-10 cycles of PCR was performed. Finally, PCR products were purified and size-selected using AMPure beads for Illumina sequencing.