pcDNA3 with Wizard

[lee shekter]

In article <1994Oct3.164057.1 at icrf.icnet.uk> g_watson at icrf.icnet.uk writes:
>I thought some fellow strugglers would like to hear my exprience using
>Promega's Wizard maxi-prep system.  I think its brilliant.  The yield is great
>and the UV trace looks wonderfull.  BUT I've tried it with Invitrogens pcDNA3
>                                    ***
>and guess what,  it does not cut :-(.
>	Well I faffed about for a while then hit on this really novel idea...
>Phenol; Phenol Cholorform IAA; Chorlorform IAA; Ether; Ethanol precipitation. 
>All this after Promega's stuff.  Well the yield was lousy (about 30% of
>starting stuff :-( ).  But it did cut  :-).
>	The odd part of this saga is that bluescript, when treated exatly
>the same as pcDNA (without the novel phenol and chloroform etc), cut perfectly
>with those same recalcitrant enzymes that would not cut the pcDNA without
>subsequent messing about.
>	I hope in the future to discover which of the above steps is the one
>that removes whatever it is that is stopping my enzymes working.  The
>experimental design is quite complex and the grant application has already gone
>in  :-)  The real question however is what the hell is causing this most
>unco-operative behaviour.  I guess it must be a protein.
>	 Anyone have any ideas or similar problems?
>		Graham Watson
>		ICRF
>		London

I've used the Wizard system on pcDNA3 and haven't had any problem with res.
enzyme cutting save for EcoRI, which doesn't appear to cut any DNA isolated by
the Wizard method.
-- 
Lee R. Shekter
Dept. of Pharmacology and Physiology         
The University of Chicago
Chicago, Illinois 60637