ChIP-Atlas: SRX5243574

Antigen

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: Cells were fixed with 1% formaldehyde for 15 minutes, thoroughly washed with DPBS, and resuspended in ChIP lysis buffer #1 (10mM Tris pH 7.4, 10mM NaCl, 0.5% NP-40, 1x HALT, 30nM Panobinostat and 5μM Nicotinamide). After sitting on ice for 10 minutes, cells were briefly vortexed and nuclei were pelleted. Nuclei were treated with MNase (NEB, M0247S) for 25 minutes at RT, pelleted and resuspended on ice in ChIP lysis buffer #2 (50mM Tris HCl pH 8.0, 10mM EDTA, 0.5% SDS, 1x HALT, 30nM Panobinostat and 5μM Nicotinamide). Chromatin was further sheared by sonication using the Sonic Dismembrator 500 (ThermoFisher Scientific) and preserved at -80oC until immunoprecipitation. 20-40ug chromatin was used for each IP 2ng of immunoprecipitated DNA from each reaction was used to create libraries using the Ovation Ultra-Low Library prep kit (Nugen, 0344-32) following manufacturer recommendations and libraries were deep sequenced on the HIseq 4000 or NextSeq 500 using single-end 50bp or single-end 75bp sequencing, respectively.