Label free analysis of nucleic acids
View Patent ↗A method is provided for label free analysis of nucleic acid materials. In accordance with the present invention, a fluorescent material is provided having a certain fluorescence emission without nucleic acids attached thereto. The fluorescent material of the present invention, has a greater emission upon the binding of a single stranded nucleic acid and an even greater emission upon the binding of a double stranded nucleic acid allowing detection of double stranded binding without using a label attached to the DNA.
1. A method for detecting nucleic acid hybridization without the use of a labeled probe, comprising:
providing a fluorescent polymeric matrix having a first fluorescent emission intensity;
attaching a single stranded DNA or RNA species to said fluorescent matrix to provide a second, greater fluorescent intensity; and
hybridizing said single stranded RNA or DNA to a homologous nucleic acid, causing a further increase in said fluorescent intensity, allowing detection of said hybridization of said homologous nucleic acids.
2. The method of claim 1 , wherein said fluorescent polymeric matrix comprises a polyvinylpyrrolidone (PVP).
3. The method of claim 1 , wherein said fluorescent polymeric matrix comprises a fluorescent film.
4. The method of claim 3 , wherein said fluorescent film is a PVP-co-poly(N-2-methacryloxyethyl)methacrylamide.
5. The method of claim 3 , wherein said fluorescent film is attached to a 3-acryloxypropryl trimethoxysilane coated surface.
6. The method of claim 3 , wherein said fluorescent film is attached to a 3-acrylamidopropyltrimethoxysilane coated surface.
7. The method of claim 4 , wherein said PVP-co-poly(N-2-methacryloxyethyl)methacrylamide is attached to a 3-acryloxypropryl trimethoxysilane coated surface.
8. The method of claim 4 , wherein said PVP-co-poly(N-2-methacryloxyethyl)methacrylamide is attached to a 3-acrylamidopropyltrimethoxysilane coated surface.
9. The method of claim 1 , wherein the fluorescent polymeric matrix is on a solid substrate.
10. The method of claim 9 , wherein the solid substrate is an array.
11. The method of claim 10 , wherein the solid substrate is glass.
12. A method for detecting nucleic acid hybridization without a labeled probe, comprising:
hybridizing a nucleic acid molecule to a single stranded RNA or DNA species attached to a solid substrate,
wherein the surface of the solid substrate comprises a fluorescent polymeric matrix and
wherein hybridizing the nucleic acid molecule to the single stranded RNA or DNA causes an increase in fluorescent activity over the fluorescent activity of the polymeric matrix and the polymeric matrix to which the single stranded RNA or DNA is attached.
13. The method of claim 12 , wherein said fluorescent polymeric matrix comprises a polyvinylpyrrolidone (PVP).
14. The method of claim 12 , wherein said fluorescent polymeric matrix comprises a fluorescent film.
15. The method of claim 14 , wherein said fluorescent film is a PVP-co-poly(N-2-methacryloxyethyl)methacrylamide.
16. The method of claim 14 , wherein said fluorescent film is attached to a 3-acryloxypropryl trimethoxysilane coated surface.
17. The method of claim 14 , wherein said fluorescent film is attached to a 3-acrylamidopropyltrimethoxysilane coated surface.
18. The method of claim 15 , wherein said PVP-co-poly(N-2-methacryloxyethyl)methacrylamide is attached to a 3-acryloxypropryl trimethoxysilane coated surface.
19. The method of claim 15 , wherein said PVP-co-poly(N-2-methacryloxyethyl)methacrylamide is attached to a 3-acrylamidopropyltrimethoxysilane coated surface.
20. The method of claim 1 , wherein the fluorescent polymeric matrix is on a solid substrate.
21. The method of claim 20 , wherein the solid substrate is an array.
22. The method of claim 21 , wherein the solid substrate is glass.