title: GSM9052521: hlsec, Vehicle, Hi-C, rep2; Homo sapiens; Hi-C
library_construction_protocol: Hi-C library preparation was carried out following existing protocols. Briefly, approximately 100,000 to 1,000,000 cell pellets were used as input per sample. Nuclei were digested overnight with MboI at 37 °C, followed by end-labeling of the DNA with biotin using 25 μl of fill-in master mix at 37 °C for 2 hours. The DNA was then ligated using 450 μl of ligation master mix at room temperature for 4 hours. DNA was purified using phenol-chloroform extraction. To remove linear DNA, the sample was treated with Lambda exonuclease (NEB) and of Exonuclease I (NEB) at 37 °C for 30 minutes. Fragmentation of 100–500 ng of DNA was performed using the TruePrep DNA Library Prep Kit V2 (Vazyme). Biotin-labeled DNA was enriched by pull-down with Dynabeads MyOne Streptavidin T1 beads (Life Technologies). Library amplification and size selection were conducted using TruePrep Amplify Enzyme (Vazyme) and SPRIselect Beads (Beckman Coulter). Libraries were sequenced on Illumina platform.