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[Yeast] Transformation problems

Margarida Palma via yeast%40net.bio.net (by mip from fct.unl.pt)
Mon Mar 5 06:50:51 EST 2007

I am having difficulties transforming an industrial strain of /_S.
cerevisiae_/  by homologous recombination in URA3. I have tried the
Gietz high-efficienty protocol and electroporation (Manivasakam and
Schiestl, 1993). I am selecting for G418 (200ug/ml). None of the
protocols had given me colonies. I am using approximately 5ug of a 5kb
DNA fragment. Is this too much?

Best regards


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