Factors affecting the effciency of end-labelling by DNA kinase

John Watson watson_j at bms.com
Tue Oct 22 17:11:40 EST 1996


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.

Well, kinasing with alpha-32P-dCTP will lead to *very low* efficiencies.  My 
advice is to follow the forward rxn protocol in Sambrook et al., the Redbook, 
etc.

AJW
----------------
John Watson
Bristol-Myers Squibb Co.
watson_j at bms.com
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