GSM7438627
In situ Hi-C experiments were conducted following the method described by Rao and colleagues with slight modifications (Rao et al., Cell. 2014). Briefly, HindIII enzyme was used to digest cells overnight at 37°C with 500U, followed by biotin filling and proximity ligation for 4 hours at 18°C with 2000U T4 DNA Ligase. After reverse-crosslinking, DNA was purified and sheared to ~300 bp fragments using Covaris S220 sonicator. The biotin-containing fragments were then immobilized on MyOne Streptavidin T1 beads. The NEXTflex adaptors were ligated, and the fragments were PCR amplified for 8 cycles using the KAPA HiFi Library Amplification Kit. Double-size selection was performed using AMPure XP beads to isolate fragments between 300 and 800bp