GSM5114279: MV411_parental_H3K27me3; Homo sapiens; ChIP-Seq
For histone ChIP-seq analysis, cells were cross-linked for 10 min with 1% formaldehyde at room temperature on a shaker at 850rpm. Crosslinked nuclei were quenched with 0.125 M glycine for 5 minutes at room temperature and washed with PBS (containing protease inhibitor and HDAC inhibitor Sodium Butyrate (NaBut)). For ChIP-seq analysis of BCOR, BCORL1, RNF2, KDM2B, and SUZ12, cells were first fixed with 2 mM DSG for 45 min at RT, followed by formaldehyde cross-linking as described above. After fixation, pellets were resuspended in 500 ul of 1% SDS (50 mM Tris-HCl pH 8, 10 mM EDTA) and sonicated in 1 ml AFA fiber millitubes for 25 minutes using a Covaris E220 instrument (setting: 75 peak incident power, 5% duty factor and 200 cycles per burst). Chromatin was immunoprecipitated using primary antibody and Dynabeads® Protein A/G. ChIP-seq libraries were made using the ThruPLEX DNA-seq 48D Rubicon kit and purified. 75-bp single-end reads were sequenced on an Illumina NextSeq instrument.