I am going to grow yeast cells in the lab for extended period of time spanning many generations. I want the population to experience haploid and diploid state in a synchronized and controlled manner. Say start from a pure diploid population, I wish to do some trick so that after overnight, the mass majority of the cells now become haploid. And vice versa.
Thanks!
Hi there,
By performing yeast mating test using a and alpha appropriate mating strain as described here:
http://www-sequence.stanford.edu/group/yeast_deletion_project/matingtest.html
Diploids won't mate (the drawback is that haploids deficient for mating won't mate either but if the purpose is just to determine ploidy of a given strain which haploid is actually able to mate, it's OK)
You could also confirm the ploidy of the population with flow cytometry.
HI,
Yes with the mating test if your new lineage mate , you can be sure that the lineage is haploid with mating type a or alpha. However what kind of trick do you plan? If you perform something enabling leading to aneuploidy then I am not so sure everything might be smooth... If nothing happens in the test cross, you cannot say that you are sure they are diploid. I would also agree with flow cytometry, and use the protocole of Haase and Reed ([Cell Cycle 1:2, 132-136, March 2002) with Sytox green which is far better than propidium iodide.
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