Hi-C of  ventricle:Embryo period 71 days female pig
The tissue was treated with 4% formaldehyde for 30 minutes for crosslinkingChromatin extraction was performed by suspending the crosslinked tissue in a lysis.The experiment continued with biotin labeling of DNA ends, which was carried out at room temperature for 45 minutes, followed by enzyme inactivation at a high temperature. DNA ligation was achieved by adding T4 DNA ligase and incubating for 4-6 hours. After ligation, proteinase K was introduced to reverse the cross-linking during an overnight incubation at 65. DNA fragments were purified, dissolved in water with unligated ends removed, and then fragmented to a size ranging from 350-500 base pairs. DNA fragments labeled with biotin were isolated using Dynabeads M-280 Streptavidin. Finally, quality control was applied to Hi-C libraries, which were subjected to sequencing on an Illumina sequencer buffer composed of 1 M Tris-HCl, 1 M NaCl, 10% CA-630, and protease inhibitors, followed by incubation on ice for 15 minutes. After crosslinking and lysis, the chromatin was digested with 400 U of Mbo restriction enzyme at room temperature.