GSM8524425: ChIP-seq mouse islets H3K27ac; Mus musculus; ChIP-Seq
~5 million cells were crosslinked in 1% formaldehyde for 10 min at RT in rotation. After quenching in 0.125 M glycine for 5 min, cells were washed in PBS and lysed in ChIP lysis buffer (2% Triton X-100, 1% SDS, 100 mM NaCl, 10 mM Tris-HCl pH 8.0, 1 mM EDTA and protease inhibitors) on ice for 30 min. Chromatin lysate was sonicated for 8 min (aiming for fragments between 150-400 bp) using a S220 Focused Ultrasonicator (Covaris) Libraries were amplified by PCR (number of cycles were determined by qPCR using the Ct + 1 value) with KAPA HiFi Hotstart Ready mix (Kapa Biosystems) and size-selected (250–350 bp) employing Agencourt AMPure XP beads (Beckman Coulter). Bioanalyzer was used for sample QC and library pooling for multiplexing.