Detection of chemical ligation using fluorescence quenching leaving groups
First Claim
1. A composition comprising a fluorophore compound, the fluorophore compound comprising a fluorophore group and a fluorescence quenching leaving group, wherein said fluorescence quenching leaving group is a dabsyl group, a dimapdabsyl group, a p-dimethylaniline-sulfonyl leaving group, a tetramethyl-phenylenediamine-sulfonyl leaving group, a nitro benzenesulfonyl group, a dinitrobenzene-sulfonyl leaving group, wherein said fluorescence quenching leaving group contains a sulfur atom bonded to three oxygen atoms and a carbon chain.
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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.
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Citations
47 Claims
- 1. A composition comprising a fluorophore compound, the fluorophore compound comprising a fluorophore group and a fluorescence quenching leaving group, wherein said fluorescence quenching leaving group is a dabsyl group, a dimapdabsyl group, a p-dimethylaniline-sulfonyl leaving group, a tetramethyl-phenylenediamine-sulfonyl leaving group, a nitro benzenesulfonyl group, a dinitrobenzene-sulfonyl leaving group, wherein said fluorescence quenching leaving group contains a sulfur atom bonded to three oxygen atoms and a carbon chain.
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35. 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 (36, 37, 38, 39, 40, 41, 42, 43, 44)
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- 46. A probe pair comprising a first and second probe, wherein said first probe comprises a fluorophore group and a fluorescence quenching leaving group, said fluorescence quenching leaving group containing a sulfur atom bonded to three oxygen atoms and a carbon chain, and selected from the group consisting of a dabsyl group, a dimapdabsyl group, a p-dimethylaniline-sulfonyl leaving group, a tetramethyl-phenylenediamine-sulfonyl leaving group, a nitro benzenesulfonyl group, a dinitrobenzene-sulfonyl leaving group, wherein said second probe comprises a nucleophilic group.
Specification