ChIP-Atlas: SRX2867686

Antigen

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: ChIP was performed as described in Gupta-Hinch et al. 2014, with modifications. Crosslinking was performed in 1% formaldehyde for 5 minutes. Input chromatin was "pre-cleared" to remove chromatin bound non-specifically by the beads. For each sample, 50 ul of equilibrated magnetic beads were resuspended in 100 ul PBS/BSA and added to the chromatin samples for pre-clearing for two hours at 4C with rotation. Beads were removed, and 100 ul of pre-cleared chromatin was set aside for the input control. 5 ul ChIP-grade rabbit polyclonal antibody was added to the remaining pre-cleared chromatin and incubated overnight at 4C with rotation. 50 ul beads were washed and resuspended as before, then incubated with the chromatin samples for two hours at 4C with rotation. After washing and decrosslinking, samples were further incubated with 80 ug RNAse A at 37C for 60 minutes and then with 80 ug Proteinase K at 55C for 90 minutes. Libraries were prepared for sequencing using standard Illumina protocols by the Oxford Genomics Centre.