mouse embryonic stem cells stably expressing a cRAF-ErT2 expression cassette
cell type
mouse embryonic stem cells stably expressing a cRAF-ErT2 expression cassette
treatment
4OHT for 2hrs
strain
E14tg2a
background strain
129SV
Stage
embryonic
antibody
input sample
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells we fixed in 1% formaldehyde at RT for 10 minutes and subsequently 2mM disuccinimidyl glutarate , nuclei extracted and chromatin sonicated using a bioruptor (10 cycles, 30s on 30s off). Cleared chromatin was incubated with anti-FLAG antibody (Sigma, F3165). Input DNA serves as negative control for antibody enrichment. Libraries were prepared using the NEB Next Ultra 2 kit.