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DSBs are repaired slowly in MCD1 simplex as compared to WT cells.
Logarithmically growing MCD1 simplex and WT strains in YPDA medium at 30°C were arrested in nocodazole for 2.5 hours and irradiated with the indicated dose (See Materials and Methods). Samples were taken before and immediately after radiation. Cells were re-suspended in warm YPDA medium containing nocadazole and incubation was continued. Samples were taken at 1 and 4 hr. (A) The chromosomal DNA was displayed using PFGE. (B) The induction of DSBs and subsequent repair was calculated from the intensity of the retained chromosomes. A detailed description of the preparation of DNA plugs, PFGE conditions and DSB quantification is presented in Materials and Methods.