Re-Transformation of BAC-plasmid-Help needed
Messerle at lmb.uni-muenchen.de
Thu Sep 19 12:43:32 EST 1996
I got a very nice BAC-plasmid of about 230 kb. For
mutagenesis I have to retransform the BAC into a
different E.coli strain. The electroporation
efficiency is very poor. Furthermore, up to now I
always ended up with much smaller plasmids after
retransformation, even when I retransformed the
plasmid into the original E.coli strain DH10B. I
isolated the BAC plasmid by alkaline lysis followed
by phenol/cloroform extraction and EtOH
precipitation. I tried to handle the DNA as gentle
possible (cut tips etc.) and expected a high
percentage of supercoiled DNA.
Any helpful suggestions?
Thanks a lot.
Max v. Pettenkofer Institute for Virology am
EMail: Messerle at lmb.uni-muenchen.de
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