GSM7727907: Tumor-derived cell line from 690108 (TOPCDS), H3K27Ac ChIP; Mus musculus; ChIP-Seq
ChIP was performed using the Active Motif ChIP-IT High Sensitivity kit (ActiveMotif, CA, USA). In brief, cells were seeded into 150mm dishes and cultured in standard growth medium. At 80% confluency, the cells were crosslinked in 37% formaldehyde (with methanol) for 15 minutes on plate and quenched with glycine.  Washed cell pellets were manually lysed using a dounce homogenizer and the chromatin was fragmented using a Q125 sonicator (Qsonica, CT, USA) with the following settings: 25% amplitude, 30 seconds 'ON'/30 seconds 'OFF' for 20 minutes total. Separate immunoprecipitation reactions using an Anti-HA tag antibody (Abcam; ab9110) or anti-Histone H3K27ac antibody (ActiveMotif, 39134) were setup for overnight immunoprecipitation reactions at 4C. ChIP DNA was bound to Protein G agarose beads, column purified, and eluted. 1-10ng of purified ChIP DNA (or input) was used to make 150bp paired-end libraries for sequencing using the TruSeq DNA library Prep Kit (Illumina, CA, USA)