C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/68 (2006.01) C07H 21/04 (2006.01) C12M 1/36 (2006.01) G01N 15/06 (2006.01)
Patent
CA 2415493
Triplex complexes contain a single-stranded probe bound to a double-stranded nucleic acid target, in which the probe includes a heteropolymeric nucleic acid or a heteropolymeric nucleic acid analog. All base triplets of the complex are members selected from the group consisting of A-T-A, T-A-T, U-A-T, T-A-U, A-U-A, U-A-U, G-C-G and C-G-C. A cation-facilitated assay includes detecting the presence of such triplex complexes to determine the degree of complementarity between the probe and target sequence. The assay preferably detects a change in fluorescent intensity of a label as a function of binding affinity between the probe and target. The label can be covalently tethered to the probe or to the target, or can be an intercalating fluorophore in the reaction medium.
Les complexes triplex contiennent une sonde ~ brin unique li~e ~ un acide nucl~ique cible ~ double brin, la sonde comprenant un acide nucl~ique h~t~ropolym~rique ou un analogue d'acide nucl~ique h~t~ropolym~rique. Tous les triplets de base du complexe sont des ~l~ments choisis dans le groupe compos~ de A-T-A, T-A-T, U-A-T, T-A-U, A-U-A, U-A-U, G-C-G et C-G-C. L'invention concerne une ~preuve facilit~e par cation qui consiste ~ d~tecter la pr~sence de tels complexes triplex afin de mesurer le degr~ de compl~mentarit~ entre la s~quence de la sonde et celle de la cible. L'~preuve permet de d~tecter, de pr~f~rence, une modification dans l'intensit~ fluorescente d'un marqueur en fonction de l'affinit~ de liaison entre la sonde et la cible. Le marqueur peut Útre li~ par covalence ~ la sonde ou ~ la cible, ou bien il peut Útre constitu~ d'un fluorophore intercalant dans le milieu de r~action.
Daksis Jasmine I.
Erikson Glen H.
Picard Pierre
Gowling Lafleur Henderson Llp
Ingeneus Corporation
Ingeneus Inc.
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