HIC DNA seq of Glyphodes pyloalis: Adult
Cells were fixed with formaldehyde to create covalent links between physically proximate DNA segments within the nucleus, thus preserving native chromatin interactions. The reaction was quenched with glycine to ensure stability of the structures.The fixed chromatin was subsequently digested with the GATC restriction enzyme. After the digestion, the DNA ends were filled in with nucleotides, incorporating biotin-14-dATP at the site of the cut for later purification. High-efficiency proximity ligation was then performed to cause covalently linked DNA fragments to ligate, resulting in chimeric molecules that represent the original chromatin architecture.Samples were sheared to a size of ~300 to 700 bp through sonication. Biotinylated DNA was pulled down using streptavidin beads to selectively enrich for ligation junctions.Library preparation involved repairing DNA ends, adding A-tails, and ligating Illumina sequencing adapters. Purified ligation products were then subjected to high-throughput sequencing using the Illumina platform to generate a large number of short-read sequences.