Applications of fluorescent n-nucleosides and fluorescent...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C07H 19/02 (2006.01) C07H 19/04 (2006.01) C07H 21/00 (2006.01) C07H 23/00 (2006.01) C12Q 1/70 (2006.01)

Patent

CA 2145750

Structural analogs of the six non-fluorescent N-nucleosides commontly found in RNA and DNA, which are inherently fluorescent under physiological conditions, are identified and methods for their preparation provided. Such analogs may be incorporated into DNA and/or RNA oligonucleotides via either enzymatic or chemical synthesis to produce fluorescent oligonucleotides having prescribed sequences. Such analogous sequences may be identical to, or the analogous complement of, template or target DNA or RNA sequences to which the fluorescent oligonucleotides can be hybridized. Methods of preparing either RNA or DNA oligonucleotide probes of the invention, intermediates used in such methods, and methods of using the probes of the invention in oligonucleotide amplification, detection, identification, and/or hybridization assays are also provided.

On a identifié des analogues structurels des six N-nucléosides non fluorescents que l'on trouve habituellement dans l'ARN et dans l'ADN, fluorescents de façon inhérente dans des conditions physiologiques et on a mis au point des procédés servant à les préparer. Ces analogues peuvent être incorporés dans des oligonucléotides d'ADN et/ou d'ARN par l'intermédiaire d'une synthèse enzymatique ou chimique, de manière à produire des oligonucléotides fluorescents possédant des séquences prescrites. Ces séquences analogues peuvent être identiques à des séquences d'ADN ou d'ARN gabarit ou cible, ou représenter le complément analogue desdites séquences, auxquelles peuvent s'hybrider les oligonucléotides fluorescents. L'invention concerne également des procédés de préparation de sondes d'oligonucléotides d'ARN ou d'ADN de l'invention, des intermédiaires utilisés dans lesdits procédés et des procédés d'utilisation des sondes décrites dans des méthodes d'amplification, de détection, d'identification et/ou d'hybridation d'oligonucléotides.

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