NOVEL METHOD OF ASSAYING NUCLEIC ACID USING LABELED NUCLEOTIDE
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Abstract
A novel method is provided to assay at least one nucleic acid. According to this method, a nucleic acid polymerization reaction is conducted in a nucleic acid polymerization reaction system, which contains (A) the at least one nucleic acid as a template, (B) at least one nucleotide monomer labeled with at least one label selected from the group consisting of (a) fluorescent dyes, (b) quenchers and (c) immune related substances with a fluorescent dye or quencher contained therein, and (C) at least one nucleic acid-synthesizing enzyme. The template nucleic acid or a nucleic acid, which has been synthesized using the template nucleic acid as a template, is then assayed from a change or an amount of a change in an optical character of the nucleic acid polymerization system. This method makes it possible to specifically and accurately assay at least one nucleic acid, which is contained in a single system and can be an unknown nucleic acid and/or a known nucleic acid, with excellent sensitivity, in short time and with ease.
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Citations
32 Claims
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1. -18. (canceled)
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19. :
- A method of assaying at least one nucleic acid, which comprises;
conducting a nucleic acid polymerization reaction in a nucleic acid polymerization reaction system comprising; (A) said at least one nucleic acid as a template, (B) (a) a fluorescent dye capable of emitting fluorescence upon binding to a nucleic acid; and
(b) a nucleotide monomer labeled with an acceptor fluorescent dye, and(C) at least one nucleic acid-synthesizing enzyme, and assaying said template nucleic acid or nucleic acid, which has been synthesized using said template nucleic acid as a template, from a change or an amount of a change in an optical character of said nucleic acid polymerization system. - View Dependent Claims (20, 21, 22, 23, 24, 25)
- A method of assaying at least one nucleic acid, which comprises;
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26. :
- A method of detecting a single nucleotide polymorphism in a nucleic acid template, the method comprising;
conducting a single-base elongation reaction in a nucleic acid polymerization reaction system, wherein the system comprises (A) the nucleic acid as a template, (B) (a) a fluorescent dye capable of emitting fluorescence upon binding to a nucleic acid; and
(b) a dideoxynucleotide monomer labeled with an acceptor fluorescent dye,(C) at least one nucleic acid-synthesizing enzyme, and (D) a nucleic acid primer, and assaying the nucleic acid template or the synthesized nucleic acid from a change or an amount of change in an optical character of the nucleic acid polymerization reaction system. - View Dependent Claims (27, 28, 29, 30, 31, 32)
- A method of detecting a single nucleotide polymorphism in a nucleic acid template, the method comprising;
Specification