cloning NdeI cut products

Amy Anne Caudy aacaudy at artsci.wustl.edu
Thu Feb 26 10:51:06 EST 1998


Richard P. Grant (spam.blocked at delete.this.cmtech.co.uk) wrote:  
<snip> 
:Alternatively, the gel purification method could be screwing up your
: ends,it has been known.  Yes, we sell a kit for doing this too (-: ,
: but have you actually tried the religation without gel purification?
: You might get high background but at least you'll be narrowing down the
: possibilities. 

	Skipping the gel purification has worked for me....also, I suggest
purification in a polyacrylimide gel.  You'll need to use quite a bit to
see it on EtBr, and of course the fragment size has to be separable.  If
you fit those parameters, it might do the trick.  Polyacrylimide is much
cleaner than agarose.  Just slice and elute in TE. 


-Amy Caudy




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