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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.


weird results after transformation

weird results after transformation - Yeast Forum

weird results after transformation - 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 04-03-2007, 04:25 PM
Margarida Isabel Rosa Bento Palma
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Default weird results after transformation



Dear all

Last time I wrote I was having problems with my yeast transformations. The
ones that answered me gave me good advices (I didn't know that I should
plate first in YPD and after replica plate in YPD+G-418) and i finally got
some colonies. Now I am having other problems. I am doing the insertion of
a fragment into S. cerevisiae chromossome (polyploid strain) by homologous
recombination. The insertion of my fragment will give this strain the
capacity of grow in G-418 because it contains the Kanamicin gene. I got
some S. cerevisiae colonies resistant to G-418 but apparently they don't
contain the insert (I have tested by PCR and Southern). Have you noticed
some similar results? Isn't weird that the strain grows without the
insert? I am sure that my initial strain doesn't grow in G-418.

Thank you

Kind regards,

Margarida

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