Plasmid miniprep from Agrobacterium

Chris Kafer ckafer at iastate.edu
Fri Jul 7 17:51:50 EST 2000


Hi.  Agro preps yield lousy and little DNA.  The way we screen is to
do a miniprep of the Agro (alkaline lysis phenol method) and then
transform  E. coli with a few microliters of this and plate.  Now you
should be able to do a regular miniprep of a culture grown from a
single E. coli colony.  If the E. coli carries the plasmid with insert
then your Agro did too!

Yes, takes an extra day or 2 but saves aggravation.

Good Luck!


On Fri, 07 Jul 2000 10:58:24 +0200, Tomek <alaska9 at friko5.onet.pl>
wrote:

>Hi,
>
>I`m lookin for efficient method of miniprep binary vector from
>Agrobacterium tumefaciens. After transformation I have very many
>colonies, but I can`t prepare plasmid pbi121 from colonies. Because my
>negative control dish is completly clear, I think, that my plasmid is
>in Agrobact. and I can`t isolate it.
>
>Thanks in advance
>
>Tomek
>
>






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