Artificial mismatch hybridization

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01)

Patent

CA 2254111

An improved nucleic acid hybridization process is provided which employs a modified oligonucleotide and improves the ability to discriminate a control nucleic acid target from a variant nucleic acid target containing a sequence variation. The modified probe contains at least one artificial mismatch relative to the control nucleic acid target in addition to any mismatch(es) arising from the sequence variation (as shown in the figure). The invention has direct and advantageous application to numerous existing hybridization methods including applications that employ, for example, the polymerase chain reaction, allele- specific nucleic acid sequencing methods, and diagnostic hybridization methods.

L'invention concerne un procédé d'hybridation moléculaire amélioré utilisant un oligonucléotide modifié qui facilite la distinction entre une cible nucléotidique témoin et une cible nucléotidique variante renfermant une variation de séquence. La sonde modifiée renferme au moins un mésappariement artificiel par rapport à la cible nucléotidique témoin, outre le ou les mésappariements qui résultent de la variation de séquence (fig. 1). Les applications aux nombreux procédés d'hybridation existants sont directes et avantageuses, par exemple l'amplification en chaîne par polymérase, les procédés de séquençage nucléotidique à spécificité allélique et les procédés d'hybridation diagnostique.

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