PCR inhibited by Redivue nucleotides???
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
It has just come to our attention, however, that Redivue
> cannot be used in PCR reactions employing Taq DNA Polymerase. The substance
> added to Redivue to stabilize the compound at higher temperatures longer is
> 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
> 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.
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