GSM9421767: PSN1-tazemetostat-Input; Homo sapiens; ChIP-Seq
Crosslinking was quenched by addition of glycine to a final concentration of 0.125 M for 5 min, followed by washing with ice-cold PBS. Cell pellets were collected and subjected to sonication to shear chromatin and centrifuged at 13,000 × g for 5 min at 4 °C. Supernatants were diluted 10-fold with ChIP dilution buffer (50 mM HEPES-KOH, pH 7.5, 500 mM NaCl, 0.5% Triton X-100, 0.5% NP-40, 0.1% deoxycholate, 1 mM EDTA). Immunoprecipitation was performed by incubating chromatin with an antibody against H3K27ac (Active Motif) for 2 h at 4 °C, followed by overnight capture with Dynabeads Protein G (Invitrogen). Beads were sequentially washed under stringent conditions, and protein–DNA crosslinks were reversed by incubation at 65 °C for 4 h. After proteinase K and RNase A digestion, immunoprecipitated DNA was purified using the QIAquick MinElute PCR Purification Kit (Qiagen). Purified DNA was used either for ChIP–qPCR validation or for ChIP–seq library preparation. Libraries were constructed according to standard Illumina protocols and sequenced on an Illumina NovaSeq X Plus platform