GSM8669406: CL246563HiC; Homo sapiens; Hi-C
Chromatin was isolated from samples using a high-throughput chromatin isolation (HIC) protocol. Formaldehyde is used to fix cells, followed by DNA digestion with the DpnII enzyme, which recognizes the GATC sequence to create 5'-overhangs. Then, dNTPs and biotin-14-dATP are utilized to fill in the 5'-overhangs, converting them into blunt ends. During this step, blunt ends are ligated, introducing a new restriction enzyme site (ClaI) into the DNA, which can be used to assess the efficiency of the filling-in reaction. After ligation, the cross-links are reversed to remove proteins from the DNA. The DNA is then cut into fragments of 200 to 300 base pairs, suitable for paired-end sequencing. Finally, streptavidin magnetic beads are employed to enrich the ligation products, and sequencing adapters are added in preparation for subsequent sequencing analysis.