GSM8112408: iNSC61, H3K27me3, ChIP; Homo sapiens; ChIP-Seq
Cells were fixed with formaldehyde-based fixing solution and lysed with provided lysis solution supplemented with protease inhibitors. Nuclei pellets were lysed, and chromatin sonicated with Bioruptor Plus sonication device (Diagenode) to obtain DNA fragments within the recommended 200–1200-bp range. In total, 25µg of sheared chromatin was then incubated with 4µg of antibody against H3K4me3 (Diagenode, C15410003-50) H3K36me3 (abcam, ab9050), H3K27ac (abcam, ab4729) or H3K27me3 (Diagenode C15410195) overnight at 4 °C with rotation. Following incubation with Protein G agarose beads, bound chromatin was washed, eluted and purified following the manufacturer's protocols. Validation by qPCR-ChIP on target genes was done before proceeding to sequencing. ChIPed DNA was end-repaired, A-tailed and adapter-ligated before size selection and amplification. The obtained libraries were QC'ed and multiplexed before 75-bp paired-end sequencing on HiSeq4000 (Illumina).