GSM8574657: HiChIP_ALL-SIL_H3K27ac; Homo sapiens; OTHER
HiChIP MNase libraries were prepared using the Dovetail HiChIP MNase Kit according to the manufacturer's protocol. Briefly, the chromatin was fixed with disuccinimidyl glutarate (DSG) and formaldehyde in the nucleus. The cross-linked chromatin was digested in situ with micrococcal nuclease (MNase) then extracted upon cell lysis. The chromatin fragments were incubated with the H3K27ac antibody (abcam – ab4729) overnight for chromatin immunoprecipitation. The antibody-protein-DNA complex was pulled down with protein A/G-coated beads. Next, the chromatin ends were repaired and ligated to a biotinylated bridge adapter followed by proximity ligation of adapter-containing ends. After proximity ligation, the crosslinks were reversed, the associated proteins were degraded, and the DNA was purified then converted into a sequencing library using Illumina-compatible adaptors. Biotin-containing fragments were isolated using streptavidin beads prior to PCR amplification. Dovetail HiChIP MNase Kit