Abstract
Random spore analysis (RSA) is a tool that allows for the screening of a large number of meiotic products. It requires only a limited effort, and is often the method of choice for constructing strains with unambiguous genotypes. It is also useful to identify the frequency of rare events. Strains are crossed on a nitrogen-limiting medium for three days. Mated cells are observed under the microscope to check for the presence of ripe asci. To release spores from their ascus, a sample of the cross is taken from the mating plate and resuspended in an enzyme solution overnight at 25–29 °C. Spores are then counted using a hemocytometer before plating an appropriate number. Incubation at the appropriate temperature follows until colonies form.
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Acknowledgments
This work was supported by NIH grant RO1 GM-81418 to S.L.F.
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Escorcia, W., Forsburg, S.L. (2018). Random Spore Analysis in Fission Yeast. In: Singleton, T. (eds) Schizosaccharomyces pombe. Methods in Molecular Biology, vol 1721. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7546-4_17
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DOI: https://doi.org/10.1007/978-1-4939-7546-4_17
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