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Origin-loading of DNA polymerase α requires Cdc45p function. Cells of strains OAy644 (WT, Polα-HA) and OAy679 (cdc45–1, Polα-HA) were synchronized in G1 phase with α factor and were released at 12°C (t = 0 min). Samples were collected at 30-min intervals (no samples collected at t = 30 or 60 min) for chromatin immunoprecipitation analysis, FACS analysis, and budding index determination (% Unbudded). PCR analysis was performed as in Fig. 1.
