HiC-Seq of Epinephelus awoara: muscle
Freshly harvested muscle tissue was cut into 2 cm pieces and immersed in a solution containing 2% formaldehyde for crosslinking. Crosslinking was halted by adding glycine, followed by additional treatment. The fixed tissue was frozen in liquid nitrogen, ground into a powder, and then resuspended in a nuclei isolation buffer to obtain nuclei. The purified nuclei were digested using HindIII and marked with biotin-14-dCTP. Non-ligated DNA ends were treated to remove biotin-14-dCTP, using T4 DNA polymerase with exonuclease activity. The ligated DNA was fragmented into 300-600 base pair (bp) pieces. The DNA fragments were repaired at the ends and given an A-tail. The DNA was then purified through biotin-streptavidin-mediated pull-down. Finally, the Hi-C library was subjected to paired-end sequencing with 150 bp read lengths using the MGISEQ-2000 platform to analyze the spatial interactions between chromosomal regions.