title: GSM8770456: H3K27ac-ChIP-seq_THP-1_SVI_0h_2025; Homo sapiens; ChIP-Seq
library_construction_protocol: ChIP-seq assays on human and mouse cells were carried out based on published protocols (Agelopoulos et al 2014; Agelopoulos et al 2012; Agelopoulos and Thanos 2006) with modifications. Cells were fixed in 1% formaldehyde (F8775, Sigma-Aldrich), for 30 minutes at room temperature, followed by quenching of the reaction by 0.125M Glycine. Cell pellets were diluted in 15 ml Buffer I [10 mM Tris-Cl pH 8.0, 10 mM EDTA, 0.25% Triton X-100] and incubated for 10 minutes on ice, followed by centrifugation at 3,500 rpm for 5 minutes at 4 °C and aspiration. Pellets were diluted in 15 ml Buffer II [10 mM Tris-Cl pH 8.0, 200 mM NaCl, 1 mM EDTA] and incubated for 10 minutes on ice, followed by centrifugation at 3,500 rpm for 5 minutes at 4 °C and aspiration. Pellets were diluted in sonication Buffer [10 mM Tris-Cl pH 8.0, 1 mM EDTA] and chromatin filaments were sheared to a range of 200-2,000 bp using a Covaris S220 Sonicator , followed by two sequential steps of 2,000 rpm and 12,000 rpm centrifugation, for 8 minutes at 4 °C, to avoid trapping of chromatin by the cell debris. Multiplexed-DNA-libraries were prepared according to a published protocol (Ford et al., 2014) with modifications. The NEBNext Multiplex Oligos for illumina (Index Primers Set 1 and 2) (E7335S and E7500S, illumina) were used to produce compatible ends to the DNA fragments.
identifiers.namespace: GEO
identifiers.id: GSM8770456_r1