Mouse ES cells routinely cultured in serum/LIF conditions
chip antibody
anti-FLAG (Sigma, catalog number F1804)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
In general, Flag ChIP followed the procedure described in Frank et al., 2001. Chromatin was fixed on 1% formaldehyde for 10 minutes and was sonicated on a bioruptor (Diagenode) to produce fragments of 100-500 bp. 500 μg of protein were used per ChIP with 10 μg of anti-Flag antibody (Sigma, catalog number F1804) and protein-bound fragments were retrieved with ProteinG-coupled magnetic Dynabeads (Invitrogen). Sequencing libraries were prepared using the NEBNext® UltraTM DNA Library Prep Kit and Multiplex Oligos (New England Biolabs) from 5 ng of DNA.