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
