GSM5930914: 22Rv1_ChIP_input; Homo sapiens; ChIP-Seq
1× 107 22Rv1 cells were cross-linked in 1% formaldehyde and quenched in 0.125 M g lycine. Cell pellets were harvested in Farnham lysis buffer [50 mM tris-HCl (pH 7.5), 150 mM NaCl, 0.5% NP-40, 5 mM EDTA, 1% Triton X-100, 1× protease inhibitor co cktail (Roche) and 1 mM PMSF] and incubated 30 min on ice to isolate nuclei. Nuclei  were suspended in SDS lysis buffer [1% SDS, 50 mM Tris-HCl, 10 mM EDTA and 1 × protease inhibitor cocktail (Roche)] and sheared to 100-500 bp by sonication. Super natants were collected and precleared 1 h with 50 μl protein G (Invitrogen). 2% of the precleared sample served as input, and the remainder was immunoprecipitated with 5 μg antibody [H3K27ac, (ab4729), Abcam] bound to 50 μl protein G beads at 4°C over night. Beads were washed 5 times with LiCl IP washing buffer [100 mM Tris-HCl (pH 7.5), 500 mM LiCl, 1% NP-40 and 1% sodium deoxycholate] and reversed cross-links  with Proteinase K overnight at 65°C. Finally, DNA was purified using a PCR Purifica tion Kit (Qiagen) and eluted in 30 μl H2O Input and precipitated DNA were sequence d using an Illumina Hiseq4000 platform.