Factors affecting the effciency of end-labelling by DNA kinase

Pamela Norton pnorton at lac.jci.tju.edu
Wed Oct 23 14:49:56 EST 1996


In article <tyr-2-2110961232030001 at news.srv.ualberta.ca>,
tyr-2 at bones.biochem.ualberta.ca (Karl Fischer) wrote:

> In article <54g99r$tn5 at enyo.uwa.edu.au>, dvoon at cyllene.uwa.edu.au (Dominic
> Voon) wrote:
> 
> >         I am going to start labelling some PCR products using DNA kinase
> > (with alpha-32P dCTP) (etc.)
> 
> Alpha labelled?? Deoxycytidine? For a kinase reaction???
> 
> This is new to me...I label fragments/oligos with gamma 32P rATP in a PNK
> reaction but have never heard of what you are proposing.
> Sigh.... ;-)

     Karl is absolutely correct. The experiment originally proposed should
lead to zero labelling. 

     Dominic, get some gamma ATP. Also, do yourself a favor and consult one
of the popular Mol. Biol. protocols books and/or a biochem. text to make
sure you understand the different ways to label "ends" of DNA.

     Pam Norton

-- 
Pamela A. Norton, Ph.D.          Assistant Professor of Medicine
Thomas Jefferson University
Philadelphia, PA 19107           p_norton at lac.jci.tju.edu



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