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Cdc7 activation is required for Clb5 origin association. cdc7-4 or CDC7-wt strains (both CLB5-MYC) were grown to log phase in YPD at 23°C, transferred for 3 h to 37°C to inactivate Cdc7 in the cdc7-4 culture, then released in fresh YPD at 23°C. Culture samples were taken at the indicated time points for FACS analysis, anti-Myc Western blotting (corrected volumes for cell equivalence), and anti-Myc chromatin immunoprecipitation.
