GSM2627271: CD4_resting_HiC; Homo sapiens; Hi-C
Cells were cross-linked with 1% formaldehyde for 10 minutes at RT. 10 X 106 cells were lysed in 10 ml lysis buffer (10 mM Tris-HCl pH8.0, 10 mM NaCl, 0.2% NP40; 10 μl protease inhibitors (Sigma)) with rotation at 4°C for 60 minutes. The cells were then treated with 400 μl 1X NEB cutsmart buffer with 0.1% SDS at 65°C for 10 minutes, followed by addition of 44 μl 10% Triton X-100 to quench SDS. Chromatin was subsequently digested with 20 Units 4 cutters (NEB). The reaction was stopped by washing the cells twice with 600 ml wash buffer. After reversing crosslink, the purified DNA were ready for the library preparation. The libraries were processed same as Hi-C protocol (Lieberman-Aiden et al., 2009, Science. 2009 Oct 9;326(5950):289-93)