The appearance of SMD is not affected by a defect in cohesion.
The temperature-sensitive mcd1-1 and apn1/2 mcd1-1 cells were grown overnight at permissive temperature (23°C) in YPDA media. The logarithmically growing cells were shifted to non-permissive temperature (37°C), nocodazole was added and cells were incubated another 3 hours to arrest them at G2/M. Cells were then treated with MMS (0.1%, 15 min) in PBS at semi-permissive temperature (30°C) followed by incubation in YPDA with nocodazole. Cells were collected at the indicated times and processed for PFGE analysis.
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