GSM8995748: HCT116 KDM5A-/-, KDM5A Mutant transfected, H3K4me3 ChIP; Homo sapiens; ChIP-Seq
Cells were crosslinked with 1% formaldehyde for 10 min at room temperature and quenched by adding 125 mM glycine for 5 minutes. Chromatin shearing was performed with a Bioruptor (Diagenode) (30 seconds ON and 30 seconds OFF at highest power for 25 cycle at 4°C). Sheared chromatin was clarified at 20,000 x g for 15 min at 4°C and the chromatin shearing efficiency verified by agarose gel electrophoresis with an observed DNA fragment length between ~250 bp and ~750 bp. Sequencing libraries were prepared with 10 ng of immunoprecipitated DNA using the KAPA HTP Library Preparation Kit (Kapa Biosystems, KR0426). After end repair, 3' A-tailing and ligation to barcoded multiplex adapters the library was PCR amplified and purified with Ampure XP beads (Beckman Coulter, A63880). Library size and distribution was checked on an Agilent Bioanalyzer, quantified on a Qubit, normalized and run on the Illumina HiSeq 2500 sequencing system (Illumina) equipped with a mid-output flow cell at a read length of 2 x 100bp.