DETECTION OF CHEMICAL LIGATION USING FLUORESCENCE QUENCHING LEAVING GROUPS
First Claim
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1. A composition comprising a fluorophore compound, the fluorophore compound comprising a fluorophore group and a fluorescence quenching leaving group.
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Abstract
Novel compounds having a fluorescence quencher as a leaving group are disclosed. Nucleic acids and other molecules containing a fluorophore and a fluorescence quencher are disclosed as an embodiment of this invention. The use of the oligonucleotides in enzyme-free oligonucleotide ligation reactions results in an increase in fluorescence when the oligonucleotide also contains a nearby fluorophore. The ligation reactions can be performed in solution, on surfaces, or in cells.
31 Citations
49 Claims
- 1. A composition comprising a fluorophore compound, the fluorophore compound comprising a fluorophore group and a fluorescence quenching leaving group.
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17. A method of detecting intramolecular chemical ligation, the method comprising:
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providing a composition comprising a fluorophore compound, wherein the compound comprises a fluorophore group, a fluorescence quenching leaving group, and a nucleophilic group;
maintaining the composition under conditions suitable for intramolecular chemical ligation; and
determining the fluorescence of the composition. - View Dependent Claims (18, 19, 20)
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21. A method of detecting intermolecular chemical ligation, the method comprising:
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providing a first composition comprising a fluorophore compound, wherein the fluorophore compound comprises a fluorophore group and a fluorescence quenching leaving group;
providing a second composition comprising a nucleophile compound, wherein the nucleophile compound comprises a nucleophilic group;
combining the first composition and the second composition to form a reaction mixture; and
determining the fluorescence of the reaction mixture. - View Dependent Claims (22, 23, 24)
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25. A method of detecting a nucleic acid sequence of interest, the method comprising:
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providing a nucleic acid molecule suspected of comprising a nucleic sequence of interest;
providing a first nucleic acid probe that hybridizes to at least a portion of the nucleic acid sequence of interest;
providing a second nucleic acid probe that hybridizes to at least a portion of the nucleic acid sequence of interest adjacent to the first nucleic acid probe;
combining the nucleic acid molecule, the first nucleic acid probe, and the second nucleic acid probe to form a mixture;
maintaining the mixture under conditions suitable for hybridization of the first nucleic acid probe and the second nucleic acid probe to the nucleic acid molecule; and
determining the fluorescence of the mixture;
wherein;
the first nucleic acid probe comprises a fluorophore group and a fluorescence quenching leaving group;
the second nucleic acid probe comprises a nucleophilic group; and
when the first nucleic acid probe and the second nucleic acid probe hybridize to the nucleic acid molecule, the nucleophilic group can displace the fluorescence quenching leaving group. - View Dependent Claims (26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39)
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40. A kit for the detection of a nucleic acid sequence of interest, the kit comprising:
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a first nucleic acid probe that hybridizes to at least a portion of the nucleic acid sequence of interest; and
a second nucleic acid probe that hybridizes to at least a portion of the nucleic acid sequence of interest adjacent to the first nucleic acid probe;
wherein;
the first nucleic acid probe comprises fluorophore group and a fluorescence quenching leaving group;
the second nucleic acid probe comprises a nucleophilic group; and
when the first nucleic acid probe and the second nucleic acid probe hybridize to a nucleic acid molecule comprising the nucleic acid sequence of interest, the nucleophilic group can displace the fluorescence quenching leaving group. - View Dependent Claims (41, 42, 43, 44, 45, 46, 47, 48, 49)
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Specification