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Whole-protein extraction from yeast was performed according to the standard procedure (Zhang et al. 2011). A600 cell equivalents of 0.12, 0.06, or 0.03 were boiled with 1 × Laemmli buffer for 5 min, loaded onto SDS-polyacrylamide gels, transferred to nitrocellulose, and probed with antibodies against Orc1 and Orc4 (Bose et al. 2004).
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