Kankyo Engineering Co., Ltd.

National Institute of Advanced Industrial Science and Technology

Other

Japan Bioindustry Association

Agency of Industrial Science and Technology

Japan As Represented by Secretary of Agency of Industrial Science and Technology

Nippon Steel & Sumikin Eco-tech Corporation

Nippon Steel Kankyo Engineering Co., Ltd.

Shinya Kurata

Takahiro Kanagawa

Ryuichiro Kurane

Yoichi Kamagata

Kazutaka Yamada

Toyokazu Yokomaku

Osamu Koyama

Kenta Furusho

Kazunori Nakamura

Masaki Torimura

Satoshi Tsuneda

Hidenori Tani

Naohiro Noda

Kohei Ichikawa

Kyoko Takatsu

[Problems] To provide a novel mixture for assaying a target nucleic acid, characterized by enabling a nucleic acid assay while: 1) requiring no step of diluting the target nucleic acid; 2) requiring no procedure of changing a probe concentration depending on a concentration of the target nucleic acid. [Means for Solving Problems] 1) A mixture which comprises one internal standard nucleic acid and two nucleic acid probes labeled with a fluorescent dye; 2) a mixture for measuring Km value which comprises one internal standard nucleic acid having a partial mutation and one nucleic acid probe labeled with a fluorescent dye; 3) a mixture which comprises one internal standard nucleic acid and one double nucleic acid probe labeled with two fluorescent dyes; and a method for assaying a nucleic acid by making use thereof.

Mixtures for assaying nucleic acid, novel method of assaying nucleic acid with the use of the same and nucleic acid probe to be used therefor

To provide a novel mixture for assaying a target nucleic acid, characterized by enabling a nucleic acid assay while: 1) requiring no step of diluting the target nucleic acid; 2) requiring no procedure of changing a probe concentration depending on a concentration of the target nucleic acid. 1) A mixture which comprises one internal standard nucleic acid and two nucleic acid probes labeled with a fluorescent dye; 2) a mixture for measuring Km value which comprises one internal standard nucleic acid having a partial mutation and one nucleic acid probe labeled with a fluorescent dye; 3) a mixture which comprises one internal standard nucleic acid and one double nucleic acid probe labeled with two fluorescent dyes; and a method for assaying a nucleic acid by making use thereof.

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.

Method of assaying nucleic acid using labeled nucleotide

A method is provided for determining a concentration of a target nucleic acid by using a nucleic acid probe labeled with a fluorescent dye. The method comprises: providing, as the probe, a nucleic acid probe capable of reducing fluorescence emission from the fluorescent dye when hybridized with the target nucleic acid; hybridizing the probe to the target nucleic acid; and measuring a decrease in fluorescence emission from the fluorescent dye after the hybridization relative to fluorescence emission from the fluorescent dye before the hybridization.

Method for determining a concentration of target nucleic acid molecules, nucleic acid probes for the method and method for analyzing data obtained by the method

A method is provided for determining a concentration of a target nucleic acid by using a nucleic acid probe labeled with a fluorescent dye. The method comprises:

Methods for monitoring PCR amplification with fluorescently labeled probes or primers for which fluorescence changes upon hybridization to a target

Nucleic acid probes are provided, each of which is formed of a single-stranded oligonucleotide which can hybridize to a target nucleic acid and is labeled with a fluorescent dye or with a fluorescent dye and a quencher substance. The nucleic acid probes can be easily designed, permit determination, polymorphous analysis or real-time quantitative PCR of nucleic acids in short time, and are not dissociated during reactions. Nucleic acid determination methods, polymorphous analysis methods and real-time quantitative PCR methods, which make use of the nucleic acid probes, are also provided.

Nucleic acid probes, method for determining concentrations of nucleic acid by using the probes, and method for analyzing data obtained by the method

A novel nucleic acid probe for nucleic acid determination includes a single-stranded nucleic acid labeled with plural fluorescent dyes containing at least one pair of fluorescent dyes to induce FRET, the pair of fluorescent dyes including a fluorescent dye (a donor dye) capable of serving as a donor dye and a fluorescent dye (an acceptor dye) capable of serving as an acceptor dye, in which the nucleic acid probe has such a base sequence and is labeled with the fluorescent dyes so that the fluorescence intensity of the acceptor dye decreases upon hybridization with a target nucleic acid. A novel nucleic acid determination method uses the probe. The probe and method can determine one or more types of target nucleic acids in an assay system in parallel using a simple apparatus.

Nucleic acid probe and novel method of assaying nucleic acid using the same

Method for determining a concentration of target nucleic acid molecules, nucleic acid probes for the method, and method for analyzing data obtained by the method

Growing community of inventors

Tokyo, Japan

Shinya Kurata