Plasmid rescue from S.cerevisiae
bundock at rulsfb.leidenuniv.nl
Wed Jan 10 17:22:40 EST 1996
I am trying to rescue a chromosomally integrated plasmid from
S.cerevisiae by plasmid rescue.Total DNA is isolated from the
transformants and digested with BglII, ligated in a large volume
(300ul),ppt, and transformed to E.coli strain MC1061 by electroporation.
I am encountering difficulties as the plasmid carries some direct
repeats, so after rescue to MC1061 (recA+) I get the same recombination
product in the MC1061 colonies.I have tried rescueing to DH5 alpha
(recA-), and still get the same recombination product.Does anybody have
experience with this and could suggest alternative recombination
defiecient E.coli strains I could try? Thanks, Paul B.
More information about the Methods