[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 

Thank you for your help.



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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