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
___________________________________________________________________
|
Rafael Maldonado | La cita ha sido
room 6160 Eccles Institute of Human Genetics |
Department of Human Genetics | retirada por respeto
University of Utah |
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