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Meiosis pp 123-132 | Cite as

Selection of G1 Phase Yeast Cells for Synchronous Meiosis and Sporulation

  • David T. StuartEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1471)

Abstract

Centrifugal elutriation is a procedure that allows the fractionation of cell populations based upon their size and shape. This allows cells in distinct cell cycle stages can be captured from an asynchronous population. The technique is particularly helpful when performing an experiment to monitor the progression of cells through the cell cycle or meiosis. Yeast sporulation like gametogenesis in other eukaryotes initiates from the G1 phase of the cell cycle. Conveniently, S. cerevisiae arrest in G1 phase when starved for nutrients and so withdrawal of nitrogen and glucose allows cells to abandon vegetative growth in G1 phase before initiating the sporulation program. This simple starvation protocol yields a partial synchronization that has been used extensively in studies of progression through meiosis and sporulation. By using centrifugal elutriation it is possible to isolate a homogeneous population of G1 phase cells and induce them to sporulate synchronously, which is beneficial for investigating progression through meiosis and sporulation. An additionally benefit of this protocol is that cell populations can be isolated based upon size and both large and small cell populations can be tested for progression through meiosis and sporulation. Here we present a protocol for purification of G1 phase diploid cells for examining synchronous progression through meiosis and sporulation.

Key words

Sporulation Meiosis Centrifugal elutriation Cell size G1-phase Yeast 

Notes

Acknowledgements

This work was supported by research grant numbers 03673 and 262070 from the Natural Sciences and Engineering Research Council of Canada.

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Department of BiochemistryUniversity of AlbertaEdmontonCanada

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