Hybridization-triggered fluorescent detection of nucleic acids
Compositions and methods for fluorescent detection of nucleic acids are provided. The compositions can be detected by fluorescence when hybridized to a nucleic acid containing a target sequence, but are non-fluorescent in the non-hybridized state. Alternatively, the fluorescence properties of the compositions change in a detectable manner upon hybridization to a nucleic acid containing a target sequence. Methods for synthesis and methods of use of the compositions are also provided.
1. A method for detecting fluorescence in a cyanine dye having a minor groove binder attached thereto, said method comprising:
providing a cyanine dye having a minor groove binder attached thereto, which is essentially nonfluorescent in the absence of double stranded nucleic acid; and
contacting said cyanine dye with double stranded nucleic acid to trigger hybridization fluorescence.
2. The method of claim 1 , wherein intercalation of said cyanine dye having a minor groove binder into double stranded nucleic acid prevents free rotation of said cyanine dye.
3. The method of claim 1 , wherein when said cyanine dye is free in solution, free bond rotation allows the cyanine dye to transit from the excited singlet state (S 1 ) to the ground state (S 0 ) in a radiationless process.
4. The method of claim 1 , wherein said cyanine dye has an electron donating group and an electron accepting group which are covalently joined to each other by a resonance linker.
5. The method of claim 1 , wherein said minor grove binder further comprises an oligonucleotide, said oligonucleotide having a 3′ end and a 5′ end.
6. The method of claim 5 , wherein said minor groove binder moiety is covalently linked to the 5′ end of the oligonucleotide.
7. The method of claim 5 , wherein the minor groove binder moiety is covalently linked to the 3′ end of the oligonucleotide.