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Yeast Forum Discuss research on yeast the model organism. Post questions and topics on the molecular biology and genetics of Saccharomyces cerevisiae and other yeast species.


Transformation problems

Transformation problems - Yeast Forum

Transformation problems - Discuss research on yeast the model organism. Post questions and topics on the molecular biology and genetics of Saccharomyces cerevisiae and other yeast species.


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Old 03-05-2007, 11:50 AM
Margarida Palma
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Default Transformation problems



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

Margarida

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