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Diploid Sporulation q

S L Forsburg nospamforsburg at salk.edu
Tue Dec 15 11:50:30 EST 1998

Ge Qiang li (gxl29 at po.cwru.edu) writes

> I need to sporulate diploids for random spore analysis, I am growing
> them on selection media, and the colonies are sitting at RT for more
> than two weeks and there is no sporulation. Any body knows what I can
> do to induce sporulation?

Assuming by "selection media" you mean EMM, they should sporulate within
a few days.  Most people however prefer ME, which is more efficient. 
Assuming you have a problem even if they are on sporulation media--

The most likely explanation is that by maintaining your diploids for an
extended period you have inadvertantly selected for a sporulation defective
strain.   pombe diploids are normally semi-induced for sporulation
so maintaining them for long periods on minimal media imposes a
strong selection for strains that can 't sporulate.  THis is why diploids
should always be maintained on YE where possible, frequently
checked for sporulation competence, and are best made fresh before use.

For a guide to working with pombe diploids, see 
For a recipe for pombe media see 

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S L Forsburg, PhD  forsburg at salk.edu
Molecular Biology and Virology Lab          
The Salk Institute, La Jolla CA 

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