I am trying to arrest budding yeast cells at 30°C and 42°C. While at 30°C after 1.5 h with 10 ug/ml Nocodazole I observe a population of metaphase-arrested cells, at 42°C I see cells at various cell cycle stages that seem to escape Nocodazole arrest. Looking through the Internet I often stumbled upon the note, that "Nocodazole may be problematic to use in budding yeast at 37°C", for example: http://tfiib.med.harvard.edu/wiki/index.php/Yeast_Cell_Cycle_Arrests
My question is: does anybody know what is behind this inefficiency?
Hi.
Is it possible that this behaviour can be attributed to cell-to-cell heterogeneity regarding aging and lifespan. Check this paper: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5349164/
It is perhaps because, at least in part, microtubules are more stable at higher temperature (and tend to depolymerise at lower temperature).
It's a well known property of Tubulin proteins.
https://www.cytoskeleton.com/faqs/tubulin
Hello, higher temperatures degrade nocodazole, resulting in reduced efficiency. You could try using benomyl instead. Hope this helps.
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