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[Yeast] weird results after transformation

Margarida Isabel Rosa Bento Palma via yeast%40net.bio.net (by mip from fct.unl.pt)
Tue Apr 3 11:25:52 EST 2007


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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