S07002_C7_D15_TC_H3K27AC
For H3K27ac, 2x10^6 fixed cells were lysed in 60ul of MAGnify Chromatin Immunoprecipitation System Lysis Buffer (Thermo Scientific) and sonicated using Bioruptor 200 (Diagenode) for 35-45min of 30s on and 30s off cycles. H3K27ac antibodies (Abcam ab4729, lots GR183922-2 (1.75ug) or GR184333-2 (1ug)) were coupled for 2h to ProteinG Dynabeads (Thermo Scientific), and used for overnight chromatin immunoprecipitation in IP buffer (1% Triton-X, 0.1% DOC, 1x TE, 1x Roche Complete Proteinase Inhibitor tablets (RCPI)). Beads were washed five times with washing buffer (50mM Hepes pH 8, 1% NP-40, 0.7% DOC, 0.5M LiCl, 1mM EDTA, and 1x RCPI) and once with TE buffer. DNA was eluted and reverse crosslinked overnight in elution buffer (10mM Tris-HCl pH 8, 1mM EDTA, 1% SDS) at 65C. DNA was purified using Qiagen MinElute PCR Purification kit, quantified by Qubit (Thermo Scientific) and submitted to library preparation and barcoding using KAPA Hyper Library preparation kit (KAPA Biosystems). Libraries were sequenced on an Illumina HiSeq2500 or a HiSeq4000.