IUBio

PCR inhibited by Redivue nucleotides???

Alexander Kraev kraev at bc.biol.ethz.ch
Wed Oct 9 04:50:46 EST 1996


In article <k.grimaldi-ya023060040810961515210001 at nntp-server.bcc.ac.uk>,
k.grimaldi at ucl.ac.uk wrote:

> I recently came across a piece which states that RediVue nucleotides
> (Amersham) inhibit taq polymerase - does any body have any experience of
> this?

<snip>
It has just come to our attention, however, that Redivue
> nucleotides
> cannot be used in PCR reactions employing Taq DNA Polymerase. The substance
> added to Redivue to stabilize the compound at higher temperatures longer is
> severely
> inhibitory to this enzyme. This fact is not divulged in the product insert
> literature. If
> your application calls for radiolabeling of nucleic acids by PCR, using
Taq DNA
> Polymerase, choose a standard, non Redivue, 32-P labeled nucleotide.

We have not used Redivue for probe labelling with Taq, but we routinely
used gamma-ATP for cycle sequencing. Up to 15 uCi in a total of 25 ul have
no ill effect on the result, although the reactions are visibly rosy coloured.


More information about the Methods mailing list

Send comments to us at biosci-help [At] net.bio.net