Characterization of checkpoint responses to DNA damage in Saccharomyces cerevisiae: basic protocols.

In spite of certain special features of its cell cycle, the yeast Saccharomyces cerevisiae has proved to be an excellent and widely used model to study eukaryotic checkpoint responses to DNA damage. This chapter primarily summarizes selected cytological methods that are useful for initial characterization of cell cycle responses. These can be useful in order to study mutants, conditions, or selected DNA damaging agents and experimental examples are given. We have also included protocols for flow-cytometric cell cycle analysis and for determination of Rad53 phosphorylation, a commonly used indicator of checkpoint activation.