GSM4974123: RM01_CP_CTCF; Macaca mulatta; ChIP-Seq
Cells were fixed by final concentration of 1% formaldehyde (Sigma-Aldrich, F1635), mixed for 10 mins at room temperature. Fixation was quenched using 2.5 M glycine at room temperature for 5 mins and immediately centrifuged at 800g for 10 mins. Supernatant was removed and frozen in liquid nitrogen and stored at −80°C until further use. Five million fixed cells were lysed for 10min, and then sonicated for 20 cycles using Bioruptor (Diagenode). After sonication, chromatin was precleared using Protein A dynabeads. CTCF primary antibody-beads added to the precleared chromatin and incubated overnight at 4°C with rotation. Beads were then washed, extract the DNA, end repair, add A, add adaptor reaction and PCR amplification, DNA products size selection were performed step by step.