ChIP-Seq with MyoD antibody 6975 in mouse M&M MEF cells transduced with MyoD
cell type
mouse embryonic fibroblasts
genotype/variation
MyoD-/-; Myf5-/-
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells were fixed in 1% formaldehyde for 11 minutes, quenched with glycine, lysed, and then sonicated to generate final DNA fragments of ~150–600 bp. The soluble chromatin was diluted 1:10 and pre-cleared with a 1:1 Protein A:G slurry for 2 hours at 4oC. Chromatin was mixed with antibody overnight at 4oC, then Protein A:G beads for 2 hours. Beads were washed and de-crosslinked overnight for ~16 hours in 1%SDS, 0.1M NaHCO3 and 70 ug Proteinase K.