pGem4 Maxi-prep problems

Simon Hansom S.Hansom at botany.uq.oz.au
Tue Aug 8 00:20:53 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.



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