RNA self-ligation. Where to get pNp from?

tfitzwater at gilead.com tfitzwater at gilead.com
Mon Nov 6 14:19:43 EST 2000


In article <8sf23b$do7$1 at nnrp1.deja.com>,
borovkov at biospud.cws.ndsu.nodak.edu wrote:


> All,
> I know it can be done with nucleotide 3',5'-bisphoshate (pNp) or
> similar chemicals, but I can't find a commercial source of these kind
> of reagents.
> Can somebody give me a hint?
> Thanks,
> Alex


>A check through our catalogues seems to confirm that these types of
>nucleotides are not available from any obvious source. It is still
>possible that a small European or Japanese company may sell them.


>It is very easy to synthesize this type of nucleotide, although I've
>always done this to synthesize [5'-32P]Cp with gamma ATP as the donor of
>the hot phosphate. (The hot pCp is used to 3'label RNA, specifically for
>chemical sequencing). There is no reason that you couldn't do the same
>reaction with cold ATP. The reaction uses Cp (or whatever Np you want)
>and polynucleotide kinase plus ATP. In labelling reactions the Cp is in
>vast excess to drive the reaction as far as possible but this wouldn't
>be as critical for your purposes.


>David F. Spencer, PhD



A recipe for preparing pCp can be found at:
gopher://iubio.bio.indiana.edu:70/0R1383?493-/Molecular
Biology/Materials+Methods

Tim Fitzwater
Principal Research Associate
Gilead Sciencesmethods at hgmp.mrc.ac.uk


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