GSM7098859: HCT116, input; Homo sapiens; ChIP-Seq
Cells were fixed in 1% formaldehyde for 10 min, followed by quenching with 125 mM glycine and two washes with phosphate-buffered saline (PBS). The cells were lysed in cell lysis buffer (5 mM Tris-HCl [pH 8.0], 85 mM KCl, 0.5% NP-40 and protease inhibitor cocktail (Nacalai Tesque)), and then centrifuged at 2,000 rpm for 3 min at 4˚C . The pellets were further lysed with nuclei lysis buffer (50 mM Tris-HCl[pH8.0], 10 mM EDTA, 1% SDS and protease inhibitor cocktail) followed by sonication using Bioruptor (Tosho Electric Co., Ltd.). ChIP-seq libraries were prepared from 500 pg of immunoprecipitated DNA fragments using KAPA Hyper Prep Kit (KAPA Biosystems).