Thiol labeling of DNA for attachment to gold surfaces
First Claim
1. A method for immobilizing a nucleic acid, containing phosphodiester and base moieties, to a surface of a gold substrate so that the base moieties are exposed and unreacted, comprising the steps of:
- (a) thiolating the nucleic acid by substituting at least one non-bridging internucleotide oxygen atom of each phosphodiester moiety in the nucleic acid with a sulfur atom; and
(b) depositing the thiolated nucleic acid on the surface of the gold substrate under conditions such that a covalent bond is formed between each sulfur atom and the gold at the surface of the substrate thereby immobilizing the nucleic acid while leaving the base moieties of the nucleic acid exposed and unreacted.
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Abstract
A method for adsorbing nucleic acids to a surface of a gold substrate for analyzing the structure of the nucleic acid includes the steps of thiolating the nucleic acid by substituting at least one non-bridging internucleotide oxygen of each phosphodiester moiety with sulfur, depositing the thiolated nucleic acid on the gold surface, and subjecting the nucleic acid to analysis for determining the atomic or molecular structure thereof. The gold surface is prepared by subjecting a gold single crystal to mechanical polishing, electropolishing, cleaning by cycles of Ar+ sputtering and annealing under vacuum until no contamination is detected by Auger electron spectroscopy, and flame annealing and quenching in methanol. Gold films deposited on atomically flat substrates, such as mica, can also be used. The thiolated DNA is deposited on the gold surface for a sufficient time for covalent bonds to form between the sulfur and the gold. Scanning tunneling microscopy, atomic force microscopy, angle-dependent x-ray photoelectron spectroscopy, and Auger electron spectroscopy are used to analyze the nucleic acid structure.
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Citations
19 Claims
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1. A method for immobilizing a nucleic acid, containing phosphodiester and base moieties, to a surface of a gold substrate so that the base moieties are exposed and unreacted, comprising the steps of:
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(a) thiolating the nucleic acid by substituting at least one non-bridging internucleotide oxygen atom of each phosphodiester moiety in the nucleic acid with a sulfur atom; and (b) depositing the thiolated nucleic acid on the surface of the gold substrate under conditions such that a covalent bond is formed between each sulfur atom and the gold at the surface of the substrate thereby immobilizing the nucleic acid while leaving the base moieties of the nucleic acid exposed and unreacted. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A method for analyzing the structure of a nucleic acid, containing phosphodiester and base moieties, immobilized on the surface of a gold substrate so that the base moieties are exposed and unreacted, comprising the steps of:
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(a) thiolating a nucleic acid by substituting at least one non-bridging internucleotide oxygen atom of each phosphodiester moiety of the nucleic acid with a sulfur atom; (b) depositing the thiolated nucleic acid on the surface of the gold substrate under conditions such that a covalent bond is formed between each sulfur atom and the gold at the surface of the substrate thereby immobilizing the nucleic acid while leaving the base moieties of the nucleic acid exposed and unreacted; and (c) subjecting the immobilized nucleic acid to analysis means and determining the atomic or molecular structure thereof. - View Dependent Claims (12, 13, 14, 15, 16, 17, 18, 19)
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Specification