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Random Spore Analysis in Fission Yeast

  • Wilber Escorcia
  • Susan L. ForsburgEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1721)

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.

Key words

Fission yeast Schizosaccharomyces pombe Meiosis Random spore analysis Strain construction Linkage analysis Plasmid recovery Lethal allele analysis Plasmid complementation 

Notes

Acknowledgments

This work was supported by NIH grant RO1 GM-81418 to S.L.F.

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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  1. 1.Molecular and Computational BiologyUniversity of Southern CaliforniaLos AngelesUSA

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