GSM8150706: HCT116_SETDB1_KO_H3K9me3_bio-rep1; Homo sapiens; ChIP-Seq
Native Chromatin immunoprecipitation was performed for both H3K9me3 and H3K14ac and were performed in biological replicates. Briefly, cells were resuspended in lysis buffer and digested with MNase to yield mononucleosomes. H3K9me3 antibody (Diagenode, C15200146) was incubated with DynabeadsG for 2 h and then with precleared chromatin for overnight at 4°C. In the case of H3K14ac, H3K14ac antibody (Millipore‐Sigma, 07‐353) was incubated with the precleared chromatin overnight and then pulled-down by incubating with DynabeadsG for 2 h. Beads were then washed twice each with low salt buffer (20 mM Tris/HCl pH 8.0, 2 mM EDTA, 150 mM NaCl, 1% Triton X-100, 0.1% SDS) and with high salt buffer (20 mM Tris/HCl pH 8.0, 2 mM EDTA, 500 mM NaCl, 1% Triton X-100, 0.1% SDS) and then followed by two rinses with TEN buffer (20 mM Tris/HCl pH 8.0, 2 mM EDTA, 150 mM NaCl). Chromatin was eluted with ChIP elution buffer (1% SDS, 100 mM NaHCO3), digested with Proteinase K, and purified with ChIP DNA Purification Kit (Active Motif). NGS libraries were prepared with 15 to 27 ng DNA using the NEBNext® Ultra™ II DNA Library Prep Kit according to the manufacturer's instructions