YPQ231202hic-Oct-23
First, chromatin is crosslinked in situ using formaldehyde to preserve spatial interactions, followed by cell lysis and restriction digestion (often with an enzyme like MboI that creates 5' overhangs). The overhangs are filled in with biotinylated nucleotides and blunt-end ligated under dilute conditions to favor proximity ligation of cross-linked fragments. After reverse crosslinking, DNA is sheared to the desired size, and biotin-labeled junctions are enriched using streptavidin beads. Sequencing adapters are then ligated, the library is PCR-amplified, purified, QC-checked, and finally sequenced.