GSM8269047: ChIRP_input; Homo sapiens; OTHER
Cells were sequentially crosslinked by 2mM DSP at room temperature for 30min with rotation and 4% formaldehyde at room temperature for 15min with rotation. 0.25M glycine was used to stop the crosslinking. Genomic DNA was fragmentated by DNase I for 10min at 37°C with rotation and sonicated by 25% amplitude for 30s. Streptavidin beads and biotin-labeled probes were added into the supernatant to precipitate RNA-gDNA complex. To elute DNA, RNase H (NEB, M0297S) was used to digest RNA and release genomic DNA. DNA library was constructed with NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB, E7645S)