GSM8279062: JN-DSRCT, assay_1, isotype, R1, ChIP-seq; Homo sapiens; ChIP-Seq
Chromatin was crosslinked for 15 minutes at room temperature (RT) by adding methanol-free formaldehyde (1% final) in the dishes culture media. Then, formaldehyde was quenched by adding a glycine solution, and cells were scraped and centrifuged before freezing for subsequent utilization. After thawing on ince, cells were resuspended in FL buffer before nuclei extraction according to the NEXSON protocol (Arrigoni et al., Nucleic Acids Res, 2016). Immunoprecipitation was performed on sheared chromatin using H3K9ac, H3K27ac, WT1 C-terminal (GTX15249), or rabbit isotype antibodies overnight and immune complexes were captured the next day using Dynabeads Protein G beads. Crosslink was reverted with Proteinase K and DNA was purified on Qiagen PCR purification columns. NebNextUltra II Library prep kit for DNA