YAP-driven tumors under Myf5-Cre or Pax7-CreER cells
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
RD cells or cells derived from YAP-driven tumors under Myf5-Cre or Pax7-CreER cells were cross-linked in 1% formaldehyde for 10 minutes at room temperature after which the reaction was stopped by addition of 0.125M glycine. Cells were lysed and harvested in ChIP buffer Chromatin was disrupted by sonication using a Diagenode Bioruptor sonicator UCD-200 to obatin fragments of average 200-500 b in size. Suitable amounts of chromatin were incubated with specific antibodies overnight. Immunoprecipitated complexes were recovered on Protein-A/G agarose beads (Pierce) or Protein G Dynabeads (Invitrogen). After extensive washes, DNA was recovered by reverse crosslinking and purification using QIAquick PCR purification kit (Qiagen) ChIP-Seq libraries were built using the NEBNext Ultra DNA Library Prep Kit for Illumina (NEB, E7370) according to manufacturer's recommendation. Libraries for each celle lines were barcoded using NEBNext multiplex oligos for Illumina (NEB, E7335) and run on a single lane of HiSeq2000 (Illumina)