GSM8374613: H3K27ac ChIP-SKO_2.1_H3K27ac; Mus musculus; ChIP-Seq
Fresh-frozen prostatic tumor tissue was pulverized (Cryoprep Pulvrizer, Covaris), resuspended in PBS + 1% formaldehyde, and incubated at room temperature for 20 minutes. Fixation was stopped by addition of 0.125 M glycine (final concentration) for 15 minutes at room temperature, then washing in ice cold PBS + EDTA-free protease inhibitor cocktail. Chromatin was isolated from biological triplicates by the addition of lysis buffer (0.1% SDS, 1% Triton X-100, 10 mM Tris-HCl (pH 7.4), 1 mM EDTA (pH 8.0), 0.1% NaDOC, 0.13 M NaCl, 1X PIC) + sonication buffer (0.25% sarkosyl, 1 mM DTT) to the samples, which were maintained on ice for 30 minutes. Lysates were sonicated (E210 Focused-ultrasonicator, Covaris) and the DNA was sheared to an average length of ~200-500 bp. ChIPseq libraries were prepared using the KAPA HTP Library Preparation Kit according to the manufacturers' protocols; ChIP-seq libraries were uniquely indexed in equimolar ratios.