Oligonucleotides containing pyrazolo[3,4-d]pyrimidines for...

Details Oligonucleotides containing pyrazolo[3,4-d]pyrimidines for... Oligonucleotides containing pyrazolo[3,4-d]pyrimidines for...

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01) C07H 21/00 (2006.01)

Patent

CA 2327547

Oligonucleotides in which one or more purine residues are substituted by pyrazolo[3,4-d]pyrimidines exhibit improved hybridization properties. Oligonucleotides containing pyrazolo[3,4-d]pyrimidine base analogues have higher melting temperatures than unsubstituted oligonucleotides of identical sequence. Thus, in assays involving hybridization of an oligonucleotide probe to a target polynucleotide sequence, higher signals are obtained. In addition, mismatch discrimination is enhanced when pyrazolo[3,4-d]pyrimidine-containing oligonucleotides are used as hybridization probes, making them useful as probes and primers for hybridization, amplification and sequencing procedures, particularly those in which single- or multiple-nucleotide mismatch discrimination is required.

Selon cette invention, les oligonucléotides dans lesquels un ou plusieurs résidus puriques sont substitués par des pyrazolo [3,4-d] pyrimidines manifestent des propriétés d'hybridation améliorées. Les oligonucléotides contenant des analogues de base des pyrazolo [3,4-d] pyrimidines ont des températures de fusion supérieures à celles des oligonucléotides non substitués appartenant à une séquence identique, ce qui permet d'obtenir des signaux plus forts dans des dosages nécessitant l'hybridation d'une sonde oligonucléotidique à une séquence cible polynucléotidique. En outre, on améliore la discrimination du défaut de complémentarité lorsque les oligonucléotides contenant des pyrazolo [3,4-d] pyrimidines servent de sondes d'hybridation, ce qui permet de les utiliser comme des sondes et des amorces destinées aux opérations d'hybridation, d'amplification et de séquençage, en particulier à celles qui demandent une discrimination du défaut de complémentarité des nucléotides uniques ou multiples.

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