GSM8014346
Samples were fixed with 1% formaldehyde (sigma,47608). Nuclear chromatin was extracted, sonicated and fragmented to 100-400bp, 5% was removed as INPUT and the remainder was used for immunoprecipitation. Primary antibodies were incubated with beads, together with fragmented chromatin, at 4°C for 6 h. Antibody-bead-protein-DNA complexes were washed sequentially with high-salt and low-salt buffers, and then the IP product was eluted. the IP product and the input were de-crosslinked with RNase A (NEB, T3018L) and Proteinase K (NEB, P8107S). The uncrosslinked input DNA and IP DNA were purified with a kit (QIAquick PCR Purification Kit, 28106). The purified Input DNA and IP DNA were constructed using the NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB, E7645S).