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
Articles from PLoS Genetics are provided here courtesy of
Public Library of Science