T/A cloning again. To Kinase or Not

Rafael Maldonado rafael at howard.genetics.utah.edu
Thu Jan 18 14:51:18 EST 1996


On Thu, 18 Jan 1996, David Micklem wrote:

> In article <Pine.SOL.3.91.960117173641.17588A-100000 at howard>, Rafael
> Maldonado <rafael at howard.genetics.utah.edu> wrote:
> 
> 
> >I don't get the point. Kinase what? PCR product, as it is, is already 
> >kinased (I mean that it has a phosphate at the 5' end).
> >
> Is it?  If you don't kinase your oligos (I don't) when exactly does your
> PCR product acquire its 5' phosphate? 

Oooops!

Sorry guys. I just didn't realize that I always digest PCR product before 
cloning.

So the answer is: PCR products don't have a P at the 5' end. But you can 
stiil cloning them if your T-vector is nos dephosphatased.


Thanks David.

Rafa


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