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Factors affecting the effciency of end-labelling by DNA kinase

daniel jacobs d.jacobs at unsw.edu.au
Tue Oct 22 02:49:58 EST 1996


In article <54g99r$tn5 at enyo.uwa.edu.au>, dvoon at cyllene.uwa.edu.au (Dominic
Voon) wrote:

> Hi all,
>         I am going to start labelling some PCR products using DNA kinase
> (with alpha-32P dCTP) and it is quite important that I get high
> effeciency. I am just wondering if anyone could just give me a quick
> run-down on the critical factors that contribute to the effeciency of this
> experiment ? such as DNA concentration, dCTP concentration etc. Any
> pointer/comment/suggestion/advice will be much appreciated. Please email
> me directly at dvoon at cyllene.uwa.edu.au.
> 
> many thanks in advance,
> dominic

Dominic I think you will need to use gamma-32P ATP if you want to
radioactively label your PCR product as it is the gamma phosphate that is
transfered.



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