title: GSM6724030: 522 DMSO_replicate 1; Homo sapiens; ChIP-Seq
library_construction_protocol: 9.7×105 cells were fixed with 0.5% formaldehyde in serum-free DMEM for 5 min at room temperature. Fixation was quenched with 1.25 M glycine. Fixed cells were washed three times with 0.5% BSA-PBS and lysed with lysis buffer (1% SDS, 10 mM EDTA, 50 mM Tris-HCl pH 8.0) containing protease inhibitor cocktail for 10 min on ice. The chromatin was sheared by Covaris Focused-ultrasonicator S220 for 10 min. The complex of anti-acetylated histone H3 lysine 27 (H3K27ac) rabbit IgG with protein G dynabeads (Invitrogen #10004D) was mixed with sheared chromatin. After incubation 4℃ overnight, the complex was washed with low salt buffer, high salt buffer, LiCl salt buffer and Tris-EDTA. The complex was incubated in 1% SDS and 0.1 M NaHCO3 buffer, followed by incubation at 65℃ for 4 h to reverse cross linking. Released DNA was purified using AMPure (Beckman #A63881). 70 pg of DNA was used for library preparation using ThruPLEX DNA-seq Kit (Takara #R400674).