Oligonucleotides labeled with a plurality of fluorophores
First Claim
1. A labeled oligonucleotide, comprising:
- an oligonucleotide sequence that hybridizes to a target polynucleotide sequence; and
at least two photometric labeling molecules attached to said oligonucleotide;
wherein said at least two photometric labeling molecules have excitation wavelengths that are within 15 nm of one another.
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Abstract
An embodiment of the invention discloses new methods for designing labeled nucleic acid probes and primers by labeling oligonucleotides with a plurality of spectrally identical or similar dyes and optionally with one or more quencher dyes. Oligonucleotides labeled in accordance with some embodiments of the invention exhibit a detectable increase in signal, for example, fluorescent signal when the labeling dyes are separated from one another. Methods for separating the dye include cleaving the labeled oligonucleotides include using enzymes that have 5′-exonuclease activity. In one embodiment nucleic acid primers of the present invention may fluoresce upon hybridization to a target sequence and incorporation into the amplification product. Nucleic acid probes and primers of the present invention have wide applications ranging from general detection of a target nucleic acid sequence to clinical diagnostics. Major advantages of the oligonucleotides including nucleic acid probes and primers of many embodiments of the present invention are their synthetic simplicity, spectral versatility and superior fluorescent signal.
45 Citations
54 Claims
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1. A labeled oligonucleotide, comprising:
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an oligonucleotide sequence that hybridizes to a target polynucleotide sequence; and
at least two photometric labeling molecules attached to said oligonucleotide;
wherein said at least two photometric labeling molecules have excitation wavelengths that are within 15 nm of one another. - 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 method of labeling an oligonucleotide, comprising:
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providing an oligonucleotide sequence that hybridizes to a target polynucleotide sequence; and
attaching at least two photometric labeling molecules to said oligonucleotide, wherein said at least two photometric labeling molecules have excitation wavelengths that are within 15 nm of one another. - View Dependent Claims (26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42)
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43. A kit for labeling an oligonucleotide, comprising:
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an oligonucleotide sequence suitable for hybridizing to a target polynucleotide sequence; and
at least two photometric labeling molecules suitable for attachment to said oligonucleotide;
wherein said at least two photometric labeling molecules have excitation wavelengths that are within 15 nm of one another. - View Dependent Claims (44, 45, 46, 47, 48, 49)
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50. A method of using a labeled oligonucleotide comprising the steps of:
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providing a labeled oligonucleotide, wherein said labeled oligonucleotide includes an oligonucleotide sequence suitable for hybridizing to a target polynucleotide sequence;
at least two photometric labeling molecules, wherein said at least two photometric labeling molecules have excitation wavelengths that are within 15 nm of one another and, wherein said photometric labeling molecules are attached to said oligonucleotide;
contacting said labeled oligonucleotide with a sample. - View Dependent Claims (51, 52, 53, 54)
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Specification