pGem4 Maxi-prep problems

Brett brett at BORCIM.WUSTL.EDU
Tue Aug 8 17:28:08 EST 1995


>I am having trouble Maxi-prepping pGem4 ( in DH5 ). I'm using the
>Alkaline-lysis method in 'Current-Protocols" 
>, repeating Sol,n I,II and III after i have obtained the DNA pellet at the end
>of the crude lysate steps. 
>I then do an RNase treatment , 2X P/C/IAA and a P.E.G. precipitation . At ther
>end of all this the plasmid won't 
>cut to completion and there is a background smear in the lane of the gel
>,which looks like digested Genomic DNA.
>I know of a few other people who have had trouble with pGem 4 but no sloutions.

If you're willing to go to all that trouble, why not just go one step further
and CsCl band it?



Brett Lindenbach
    
Program in Immunology                              
Washington University - St Louis                  
brett at borcim.wustl.edu                             






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