GSM8256969: Dnmt1tet/tet mESCs, Dnmt1 control (CTL), replicate 2, H3K27ac ChIP-seq; Mus musculus; ChIP-Seq
Chromatin immunoprecipitation (ChIP) sequencing was performed in duplicates for each histone modification per experimental condition and one aliquot of non-immunoprecipitated chromatin (i.e. input) was sequenced per experimental condition. First, cells were fixed with a solution consisting of 13% formaldehyde (Sigma-Aldrich, 47608) in DPBS (Corning, 21-031-CV) for 10 minutes at room temperature with gentle rocking. Then, cell and nuclei lysis was performed, and chromatin was purified and sonicated using buffers and protocols from the IDeal ChIP-Seq Histone Kit (Diagenode, C01010051). Sonication was conducted with the Bioruptor® Pico sonication device (Diagenode, B01060010) for 15 cycles (30 seconds ON/ 30 seconds OFF) to obtain DNA fragments between 150-400bp. Next, chromatin was pre-cleared via incubation with protein G Dynabeads™ (Thermo Fisher, 10009D) for 1h at 4°