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Quantitating Radioactive label in DNA and RNA

Bernard Murray bernard at elsie.nci.nih.gov
Sat Jul 2 16:38:59 EST 1994


In article <aquilla.1123491816E at sadye.emba.uvm.edu>, aquilla at salus.med.uvm.edu (Tracy Aquilla) writes:
> In Article <2us9ns$eso at matt.ksu.ksu.edu>, hlhinkel at ksu.ksu.edu (Heidi Lanora
> Hinkel) wrote:
> >        I am looking for methods with which to quanitate the
> >amount of P32 -ATP incorperated by end-labeling with T4 kinase,
> >in both DNA and RNA probes. I would like to hear of any methods
> >that you know of and which one you think is best. Thanks in
> >advance
> >                Heidi Hinkel
> >                hlhinkel at matt.ksu.ksu.edu
> >                Division of Biology
> >                Kansas State University
> >                Manhattan, KS 66506
> >
> >
>  I usually just count a small fraction of the probe after purification,
> using a scintillation counter. How much more quantitative do you need to get?
> Tracy

....or does Heidi mean *specific* incorporation (dpm/ng rather than dpm/ul)?
The former is a good deal more difficult to do as the amounts of NA probe are
usually rather small (as well as being hot!).  If this *is* the case then
I can only suggest attempting to quantify the DNA (eg. by the ethidium bromide
on Saran wrap/gel method) and then counting the same spot.  I also have a
very vague recollection of being able to do this by TLC (whilst checking the
purity of the probe) but I can't remember any details.
		Good luck!
			Bernard

Bernard Murray, Ph.D.
bernard at elsie.nci.nih.gov (National Cancer Institute, NIH, Bethesda MD, USA)



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