Ti plasmid isolation from A.t...

Dr S T May lsrei at csv.warwick.ac.uk
Fri Sep 16 12:55:52 EST 1994


Hi there, 

I've used alkaline lysis to extract plasmid DNA from agro,
although I know the protocol as the Modified Burke/Ishorowitz
method (also sometimes referred to as 'the one in Maniatis :)' )

The DNA you get out is fine for re-transformation into E. coli for
insert analysis, but it is not good quality DNA and is hard
(often impossible) to restriction digest directly (and get any 
meaningful results).

I am currently using agro-colony PCR to check inserts, but if 
anyone in this forum knows of a way to reliably extract good 
quality 'cuttable' agro plasmid DNA, then please post the method
(or a pointer to it) as I know that many people would be interested
and grateful.

Thanks,
Sean 
 



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