>Hi,
>>Has anyone ever had problems growing yeast (with 2 selectable plasmids, 2u
>ori) in minimal medium? My yeast grows well on plates and really slow
>(double time approx. O/N) in liquid medium (growth stops after 2 days at
>approx. OD600=1). Apart from the yeast getting dizzy in the shaker, there
>shouldn't be any difference btwn liquid cultures and growth on solid medium.
>My LIQUID Bgal assays are not working out...HELP!
>>The shaker is at 30C and 225rpm.
>>Thanks for being there,
>>Kath
Hi Kath,
It sounds like the medium you are using is limited for some nutrient. What
MM are you using-yeast nitrogen base /wickerhams defined medium or other.
Also what yeast? If it is Saccharomyces here are
a couple of suggestions.
1. Are you sure that you know all of the auxotrophic requirements?
2. Used double /triple strength medium with 2% carbon source. If you get
increased biomass and growth rate something is definately limiting in the
original medium.
3.Try adding 0.1%casamino acids (final conc). This should supplement any
unknown auxotrophies. If you plasmids have URA markers the medium will
still be selective.
4. Buffering your medium with something like citrate phosphate buffer (
25mM) or 0.1 M succininc acid can help. This is very important if the
cultures are continuous. They will appear to grow OK in batch but will
washout when pumps are switched on.
hope this is of some help
Regards
Ronan
******************************************************************************
Ronan O'Kennedy
Growth and Fermenatation Technology Lab
Dept of Microbiology
University College Galway
Ireland
Phone ***-91-24411ext 2191
Fax ***-91-25700
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