Fibroblast cells transfected with vectors carrying MyoD gene
transduced gene
human MyoD
cell type
Fibroblast cells
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
cells were fixed with 1% formaldehyde (wt/vol) for 15 min, washed with 1 X PBS and lysed in buffer with 50 mM Tris(pH8), 1% SDS (wt/vol), and 10 mM EDTA. Lysed cells were sonicated with 30 s on/off cycles at high intensity by using a bioruptor (Diagenode). Sonicated supernatants were diluted with buffer containing 16.7 mM Tris-HCl(pH8), 0.01% SDS, 1.1% Triton X-100, 1.2 mM EDTA and 167 mM NaCl. 10 ul of FLAG antibody (Monoclonal anti-FLAG M1 antibody produced in mouse, F3040, Sigma-Aldrich) was added into the diluted supernatants, and 60 ul of Dynabeads protein A beads were added and incubated for 3 hours at 4C. ChIP-seq libraries were made by using NEBNext Ultra DNA Library prep kit for Illumina