Sma I religation problem

Barry Moore barrym at stella.med.utah.edu
Thu Jul 20 13:58:08 EST 1995


In article <1995Jul20.115013 at ania.path.ox.ac.uk> rpgrant at molbiol.ox.ac.uk (Mad Dan Eccles) writes:

   In article <3ujkm8$6h1 at hydra.acs.ttu.edu>, Amadou Ba <PHYASB> writes:

   > If we understand correctly, adding Klenow and dNTP will make 
   > every thing blunt ended.
   > 

>  Should still work, but you need to phosphatase the vector.

I would add that you want to be careful with the CIP.  I just spent the last month figuring out that
I was using to much CIP, and it was ruining my cloning (I don't know if this enzyme is so harsh that
it was chewing up my vector ends, or if it was making it through my gel purification to destroy my ligation).
I found that 0.1 U/0.5-1.0 ug vector prevent recircularization.

Barry
--

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*                  Barry Moore                   *
*               Univeristy of Utah               *
*           Department of Human Genetics         *
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*                     USA                        *
*                (801) 281-4636                  *
*            barrym at corona.med.utah.edu          *
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