title: ChIP-seq of homo sapiens: HDGF rep 2
design: ChIP was performed using a SimpleChIP Plus Enzymatic Chromatin IP Kit (9005S, Cell Signaling Technology, Danvers, MA, USA). Briefly, cells (5  106 per ChIP) were crosslinked in culture medium containing 1% formaldehyde at room temperature for 10 min, and the reaction was stopped by adding glycine solution. Cells were then lysed, and nuclei were collected and treated with micrococcal nuclease (1 L per ChIP) for 20 min at 37 C. Following termination of the reaction with 0.05 M EGTA, samples were sonicated with several pulses to disrupt nuclear membranes. Next, the supernatants, containing chromatin, were collected after centrifugation. Chromatin solutions were then incubated with HDGF, and H3K36me3 antibodies overnight at 4 C with rotation, followed by incubation with Protein G magnetic beads for 2 h at 4 C. Beads were washed, and chromatin was eluted. Crosslinks were reverted according to the manufacturers instructions. DNA was subsequently purified and used for ChIP-seq analysis.