title: GSM9080139: H3K27me3_DKO_rep_1; Homo sapiens; ChIP-Seq
library_name: GSM9080139
library_construction_protocol: Cells were cross-linked with 1% formaldehyde in PBS for 15 min at room temperature, then quenched with 125 mM glycine. Cells were scraped, pelleted at 1,000 × g for 5 min at 4°C, and stored at –80°C. Protein G magnetic beads (40 µL/sample) were washed and incubated overnight at 4°C with 0.5% BSA/PBS and 1.5 µL anti-H3K27ac antibody. Cell pellets were lysed in buffer (1 mM Tris-HCl, pH 8.0, 100 mM NaCl, 1 mM EDTA, 1 mM EGTA, 0.1% Na-deoxycholate, 0.5% N-lauroylsarcosine) with protease inhibitors, and chromatin was sonicated (Handy Sonic UR-21P) to 150–450 bp. Beads were washed with ChIP dilution buffer and incubated overnight with chromatin at 4°C. Beads were washed with low salt, high salt, LiCl, and TE + NaCl buffers, then eluted in 1% SDS buffer at 65°C for 15 min. Cross-links were reversed at 65°C for at least 6 hours. Samples were treated with RNase A (0.15 mg/mL) at 37°C for 30 min and proteinase K (0.1 mg/mL) at 55°C for 1 h. DNA was purified by phenol-chloroform extraction and ethanol precipitation. ChIP DNA libraries were prepared with the NEBNext ChIP-Seq Library Prep Kit for Illumina (NEB)
identifiers.namespace: GEO
identifiers.id: GSM9080139_r1