[Yeast] Yeast plasmids into E. coli

Anna Heidenblut via yeast%40net.bio.net (by anna.heidenblut At gmx.de)
Thu Nov 23 09:23:57 EST 2006


Dear all,

after a library screen I have extracted yeast plasmid DNA from my 
candidate clones and I tried to transform electrocompetent E. coli Top10 
with the plasmids I got out of the yeast clones. No matter what kind of 
plasmid preparation I did (I tried the Pierce DNA extraction kit and the 
Q-BIOgene plasmid DNA extraction kit) I only get E. coli colonies from 
about 50% of my DNA samples. The plasmids I'm trying to transform in 
might be rather large (10.6 kb vector plus 5 to 20 kb insert). Does 
anybody know how I could get E. coli colonies from the rest of my DNA 
samples?

Thank you for your help.

Regards,

Anna

-- 
Dr. Anna Heidenblut (PhD)

Genome Integrity Group
Department of Molecular Oncology
Weatherall Institute of Molecular Medicine
Oxford University John Radcliffe Hospital,
Headley Way, Oxford, Oxfordshire OX3 9DS UK
Tel: 0044 / 1865 / 222419
anna.heidenblut At cancer.org.uk



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