[Yeast] problem recovering transformed glycerol stock

Isaac Garcia-Murillas via yeast%40net.bio.net (by Isaac.Garcia-Murillas from icr.ac.uk)
Mon Aug 11 05:55:52 EST 2008


Hi all

I am currently working with the yeast strain YBZ-1 and I have
transformed it with several pACT2 -based plasmids (each one
individually). The transformations went fine and I had colonies only on
the plates selecting for the marker on the plasmid (LEU). I have
prepared glycerol stocks (15% and also 20%) and stored them at -80C.
Now, when I try to recover theses stocks in minimal plates the cells do
not grow, but when I recoverd them on complete media they do grow nicely
after a couple of days. If I replate them from the complete media plates
onto minimal plates they do grow, but this is time consuming as it
requieres two plate changes. They also recover nicely on liquid minimal
media, but I haven't been able to recover them on minimal plates not a
single time. The parent, untransformed, strain YBZ-1 has been frozen at
-80C in 20% glycerol and recovers very nicely on complete media plates.
I am wondering if somebody knows what is going on here, and why my
transformed yeast do not want to grow from frozen onto minimal media
plates.

Thanks a lot for your help

isaac

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