GSM4792288: H3K4me3 Hi-CHIP in USP22 wild type (shLuc), untreated cells, replicate 1; Homo sapiens; OTHER
HCT116 cells were fixed in 1% formaldehyde for 10 minutes at room temperature, followed by quenching with 0.125 M Glycine for 5 minutes. Cells were harvested and crosslinked chromatin was extracted, MboI-digested, biotin-labeled, and ligated as described. Chromatin was sheared to 100-300 bp fragments using the Q800R2 Sonicator with circulating chiller (QSonica), for 25 minutes at 50% amplitude, 20 sec on/20 sec off. Sheared chromatin was incubated with anti-H3K4me3 (Sigma #: 07-473) for 2 hours at 4°C against indicated proteins followed by wash, elution, reverse-crosslinking, biotin purification, and Tn5 tagmentation steps. Libraries were constructed using Nextera XT Index Kit (Illumina) and size selected using Ampure XP beads (Beckman-Coulter).