Kits and reaction mixtures containing modified probe molecules
First Claim
1. A kit comprising:
- a probe molecule for use in determining the presence of an RNA target in a sample, the probe comprising complementary first and second base regions that form a hybrid containing at least one 2′
-O-alkyl ribonucleotide, wherein the probe forms a stable complex with the RNA target but not with a non-targeted nucleic acid under nucleic acid assay conditions, such that the RNA target can be detected, and wherein the complex comprises a single-stranded form of the probe; and
a solid support for immobilizing the RNA target so that unbound nucleic acids and other components of the sample can be removed from the RNA target.
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Abstract
The present invention concerns oligonucleotides containing one or more modified nucleotides which increase the binding affinity of the oligonucleotides to target nucleic acids having a complementary nucleotide base sequence. These modified oligonucleotides hybridize to the target sequence at a faster rate than unmodified oligonucleotides having an identical nucleotide base sequence. Such modified oligonucleotides include oligonucleotides containing at least one 2′-O-methylribofuranosyl moiety joined to a nitrogenous base. Oligonucleotides can be modified in accordance with the present invention to preferentially bind RNA targets. The present invention also concerns methods of using these modified oligonucleotides and kits containing the same.
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Citations
47 Claims
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1. A kit comprising:
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a probe molecule for use in determining the presence of an RNA target in a sample, the probe comprising complementary first and second base regions that form a hybrid containing at least one 2′
-O-alkyl ribonucleotide, wherein the probe forms a stable complex with the RNA target but not with a non-targeted nucleic acid under nucleic acid assay conditions, such that the RNA target can be detected, and wherein the complex comprises a single-stranded form of the probe; anda solid support for immobilizing the RNA target so that unbound nucleic acids and other components of the sample can be removed from the RNA target. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24)
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25. A reaction mixture comprising:
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one or more amplification oligonucleotides in the presence of at least one nucleic acid polymerase and nucleotide triphosphates sufficient to form an RNA amplification product; and a probe molecule comprising first and second base regions hybridized to each other and having at least one 2′
-O-alkyl ribonucleotide, wherein the probe forms a stable and detectable complex with the amplification product but not with non-target nucleic acid, and wherein the complex comprises a single-stranded form of the probe. - View Dependent Claims (26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47)
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Specification