Claims
- 1. A method for extending a nucleic acid bound to a nanoparticle, the method comprising:
binding to a nanoparticle a single-stranded DNA primer; annealing to the nanoparticle-bound primer a single-stranded DNA; and enzymatically extending the primer, thereby extending a nucleic acid bound to a nanoparticle.
- 2. The method of claim 1, wherein the nanoparticle comprises one or more metals.
- 3. The method of claim 2, wherein the one or more metals is selected from gold, silver, copper, nickel, rhodium, palladium, and platinum.
- 4. The method of claim 1, wherein the primer is bound to the nanoparticle via a 5′ thiol linker.
- 5. A method for reverse transcribing mRNA directly onto a nanoparticle, the method comprising:
binding to a nanoparticle a single-stranded DNA primer; annealing to the nanoparticle-bound primer a single-stranded mRNA; and reverse transcribing the mRNA, thereby reverse transcribing mRNA directly onto a nanoparticle.
- 6. The method of claim 5, wherein the primer is a poly-dT primer.
- 7. The method of claim 5, wherein the nanoparticle comprises one or more metals.
- 8. The method of claim 7, wherein the one or more metals is selected from gold, silver, copper, nickel, rhodium, palladium, and platinum.
- 9. The method of claim 5, wherein the primer is bound to the nanoparticle via a 5′ thiol linker.
- 10. A method for determining the identity of a specific nucleotide at a defined site in a nucleic acid, the method comprising:
binding to a nanoparticle a single-stranded DNA primer via its 5′ end; annealing to the nanoparticle-bound primer a single-stranded DNA having a specific nucleotide whose identity is to be determined such that the 3′ end of the primer anneals to a nucleotide flanking the specific nucleotide whose identity is to be determined; subjecting the nanoparticle-bound primer and annealed DNA to a polymerizing agent in a mixture containing each of ddATP, ddGTP, ddCTP, and ddTTP, wherein each of ddATP, ddGTP, ddCTP, and ddTTP are labeled with a different label, such that the primer is extended by a single nucleotide; and detecting the identity of the single nucleotide added to the 3′ end of the primer, thereby determining the identity of a specific nucleotide at a defined site in a nucleic acid.
- 11. The method of claim 10, wherein the nanoparticle comprises one or more metals.
- 12. The method of claim 11, wherein the one or more metals is selected from gold, silver, copper, nickel, rhodium, palladium, and platinum.
- 13. The method of claim 10, wherein the primer is bound to the nanoparticle via a 5′ thiol linker.
- 14. The method of claim 10, wherein the polymerizing agent is a DNA polymerase.
- 15. A method for introducing sidedness to a metal particle, the method comprising:
binding to a nanoparticle a plurality of first single-stranded DNA molecules; binding to a solid support a plurality of second single-stranded DNA molecules, wherein the first and second single-stranded DNA molecules are complementary to each other; contacting the nanoparticle with the solid support such that those first single-stranded DNA molecules nearest the solid support anneal to the second single-stranded DNA molecules contained thereon, and those first single-stranded DNA molecules furthest from the solid support do not anneal to the second single-stranded DNA molecules contained thereon and thus remain free, resulting in a nanoparticle having first single-stranded DNA molecules that are unannealed and free, and first single-stranded DNA molecules that are annealed and not free; subjecting the nanoparticle to an agent that modifies those first single-stranded DNA molecules that are unannealed and free, but does not modify those first single-stranded DNA molecules that are annealed and not free; and separating the nanoparticle from the solid support, thereby resulting in a nanoparticle having first and second sides, wherein the first side contains modified first single-stranded DNA molecules, and wherein the second side contains unmodified first single-stranded DNA molecules, thereby introducing sidedness to a nanoparticle.
- 16. The method of claim 15, wherein the nanoparticle comprises one or more metals.
- 17. The method of claim 16, wherein the one or more metals is selected from gold, silver, copper, nickel, rhodium, palladium, and platinum.
- 18. The method of claim 15, wherein each of the first single-stranded DNA molecules are bound to the nanoparticle via a thiol linker.
- 19. The method of claim 15, wherein the agent is an enzyme.
- 20. A method for generating covalently immobilized DNA, the method comprising:
binding a first single-stranded DNA primer to a nanoparticle; mixing the nanoparticle with a DNA having first and second complementary strands under conditions such that the first complementary strand of the DNA anneals to the nanoparticle-bound primer; and enzymatically extending the first primer, thereby generating covalently immobilized DNA.
- 21. The method of claim 20, wherein in the mixing step, a second single-stranded DNA primer is mixed with the nanoparticle and the DNA under conditions such that the second complementary strand of the DNA anneals to the second primer, and wherein in the extending step, the second primer is enzymatically extended.
- 22. The method of claim 21, wherein the mixing and extending steps are repeated one or more times.
- 23. The method of claim 20, wherein the nanoparticle comprises one or more metals.
- 24. The method of claim 23, wherein the one or more metals is selected from the group consisting of gold, silver, copper, nickel, rhodium, palladium, and platinum.
- 25. The method of claim 20, wherein the first single-stranded DNA primer is bound to the nanoparticle via a 5′ thiol linker.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority under 35 U.S.C. § 119(e) to to U.S. Provisional Patent Application No. 60/335,151, filed Nov. 1, 2001, the disclsoure of which is herein incorporated by reference.
GRANT REFERENCE
[0002] Work for this invention was funded in part by a grant from United States National Institutes of Health Grant No. R01 HG02228, and in part by a grant froom National Science Foundation Grant No. DBI 9872629. The United States Government may have certain rights in this invention.
Provisional Applications (1)
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Number |
Date |
Country |
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60335151 |
Nov 2001 |
US |