ChIP sequencing of mus musculus: B16F10 cell lines
20 million B16F10 cells growing under GLU or GAL conditions were fixed in 1.1% PFA solution for 15 minutes. To quench the cross-linking, a final concentration of .125 M glycine was added for 5 minutes before cells were harvested. The cells were washed in chilled PBS with .5% Igepal twice, before adding PBS-Igepal with 1mM PMSF. The ChIP DNA samples were then submitted to Active Motif for preparation of libraries, sequencing, and analysis.According to Active Motif protocol, cells were lysed, sonicated, and then DNA-protein complexes were isolated with antibodies. Samples were reverse-crosslinked and DNA was purified.