Hi-C Analysis for Anisodus tanguticus: Old and Young Plants
The main process of HiC library construction is as follows: (1) Cell cross-linking: xxx (samples) were fixed with 1% formaldehyde solution in MS buffer (10 mM potassium phosphate, pH 7.0; 50 mM sodium chloride; 0.1 M sucrose) for 30 min under vacuum at room temperature. After fixation, xxx (samples) were incubated in MC buffer containing 0.15 M glycine for 5 min at room temperature under vacuum. The fixed tissue was submerged in liquid nitrogen, then resuspended in nucleus isolation buffer, filtered through a 40 nm cell strainer, and nuclei were collected. (2) Restriction enzyme digestion: The cells were lysed, and high-quality DNA was obtained. Then the cross-linked DNAwas digested with HindIII restriction enzyme. (3) Biotin labeling: the sticky end is repaired, and biotin is added at the same time to label the oligonucleotide end. (4) Cyclization: T4 DNA ligase (NEB) was added and incubated at 16 for 4~6 hours for DNAligation. (5) DNA purification: Add proteinase K to uncrosslink during overnight incubation at 65 . Then, the unligated ends were removed, the purified DNA was fragmented to a size of 300-500 bp, and the DNA ends are repaired. The labeled DNA fragment of biotin were finally isolated on Dynabeads M-280 Streptavidin (Life Technologies). (6) DNA sequencing: The obtained DNA fragments were used to construct library.