GSM8409486: NHEM_SS18_ChIP; Homo sapiens; ChIP-Seq
For ChIP-seq: For histone modifications, 10-15M cells were crosslinked with 1% Formaldehyde; For chromatin remodelers and TFs, 10-40M cells were double cross-linked with 0.25 M disuccinimidyl glutarate (DSG), followed by 1% formaldehyde. Nuclei were isolated and lysed, and chromatin was fragmented by sonication. Extracts were incubated with specific antibodies O/N, followed by 2h incubation with magnetic Protein A+G beads. After extensive washes, DNA was eluted from the beads and reverse crosslinked. For ChIC: Chromatin Immuno Cleavage (ChIC or CUT&RUN) was performed using CUTANA ChIC/CUT&RUN kit following manufacturer's protocol. Libraries for ChIP-seq were prepared according to Carcamo et al., Cell Rep 39, 110637. 10.1016/j.celrep.2022.110637. Libraries for ChIC were prepared using NEBNext Ultra II DNA library kit, following manufacturer's recommendations. ChIP-seq CUT&Run (ChIC)