GSM8831534: 436_siNT_H3K27me3_Rep1; Homo sapiens; ChIP-Seq
For ChIP, 20 million cells mixed with NIH 3T3 cells (5%) as a spike-in control and then crosslinked with 1% formaldehyde for 20 minutes. Sonicated chromatin extracts were incubated overnight with H3K27me3 antibody (Active Motif, 39155, 5 µL/IP). Protein A Sepharose beads (Cytiva, 17078001, rabbit antibodies) were subsequently added for immunoprecipitation.The immunoprecipitated chromatin was washed sequentially with high-salt buffers and TE buffer. After washing, chromatin was reverse crosslinked, and DNA was extracted. ChIP libraries were prepared using the MicroPlex Library Preparation Kit v3 (Diagenode, C05010001), following the manufacturer's protocol.