GSM7421569: C2_input; Mus musculus; ChIP-Seq
Cells were fixed with 1% formaldehyde for 10 minutes, followed by quenching with 125 mM glycine for 5 min at room temperature. Chromatin was sonicated in the Diagenode bioruptor (Diagenode Inc.). The cleared supernatant was incubated with anti-H3K27ac antibody. After adding 15 𝜇l of prewashed protein G-magnetic beads, the reactions were incubated for 3 hours. The beads were extensively washed with ChIP buffer, high salt buffer, LiCl2 buffer, and TE buffer. Bound chromatins were eluted and reverse-crosslinked at 65°C overnight. DNAs were purified using Min-Elute PCR purification kit (Qiagen) after treatment with RNase A and proteinase K. ChIP-seq libraries were prepared from 10 ng of ChIP and input DNAs with the ThruPLEX® DNA-seq Kit (Takara Bio, Cat# R400675).