Q: Klenow and CIAP simultaneously in one pot?

frosty at pai.ku.dk frosty at pai.ku.dk
Mon Jan 27 10:32:17 EST 1997


> 
> From: Wolfgang Schechinger <wgschech at med.uni-tuebingen.de>
> 
> Subject: Q: Klenow and CIAP simultaneously in one pot?
> 
> Date: 17 Jan 1997 11:43:46 -0000
> 
> Is this possible?
> 
> Any comments are appreciated!

I used to do this, but found that CIAP (at least the stuff
I buy from Boehringer) tends to damage the ends of the DNA
unless used in its own buffer.  My cloning efficiency improved
dramatically when I stopped cutting this corner.  Klenow on the
other hand works very well in most restriction endonuclease 
buffers. You can simply add dNTPs and Klenow for the last
five minutes of the digestion.

Frosty Loechel
> Wolfgang
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