RNA self-ligation. Where to get pNp from?

David F. Spencer dspencer at is.dal.ca
Fri Nov 3 15:18:00 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.

Dave

-- 
David F. Spencer, PhD
Dept. of Biochemistry and Molecular Biology
Dalhousie University
Halifax, Nova Scotia, Canada

dspencer at is.dal.ca






More information about the Methods mailing list