SINGLE MOLECULE REAL TIME SEQUENCER, NUCLEIC ACID ANALYZER AND SINGLE MOLECULE REAL TIME SEQUENCING METHOD
First Claim
1. A single molecule real time sequencer comprising:
- a nucleic acid probe having a photodegradable substance that inhibits a nucleic acid extension reaction;
a reaction field area arranged with a plurality of said nucleic acid probes;
a channel installed with a plurality of said reaction field areas;
a reagent supply mechanism for supplying a reagent to be utilized in the nucleic acid extension reaction to the channel;
an excited optical system for irradiating evanescent light to the desired reaction field area and detecting fluorescence generated; and
a reaction control optical system for irradiating the light to decompose the photodegradable substance to the desired reaction field area.
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Accused Products
Abstract
An object of the present invention relates to selectively control an extension reaction within a desired area in a substrate. In the present invention, an oligo probe arranged with a caged compound at the terminal thereof is immobilized to a reaction field area in the substrate. After pouring a reaction solution into a flow cell including the reaction field area, the reaction field area alone is irradiated with light to associate the photodegradation-active protecting group at the terminal of the oligo probe that has been immobilized in the reaction field area and thus to selectively control initiation of a polymerase extension reaction. In the flow cell, a plural number of the reaction field areas are arranged at constant interval on the substrate. The flow cell immobilized to a moving stage is moved by a distance equal to the interval between the adjacent reaction field areas and then light irradiation is carried out to measure the extension reaction continuously. By repeating these operations, the base extension reaction is stably measured in the flow cell without using complicated channels or without replacing the reaction solution.
6 Citations
23 Claims
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1. A single molecule real time sequencer comprising:
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a nucleic acid probe having a photodegradable substance that inhibits a nucleic acid extension reaction;
a reaction field area arranged with a plurality of said nucleic acid probes;a channel installed with a plurality of said reaction field areas; a reagent supply mechanism for supplying a reagent to be utilized in the nucleic acid extension reaction to the channel; an excited optical system for irradiating evanescent light to the desired reaction field area and detecting fluorescence generated; and a reaction control optical system for irradiating the light to decompose the photodegradable substance to the desired reaction field area. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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9. A nucleic acid analyzer comprising:
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a nucleic acid analysis device having a nucleic acid probe that hybridizes with a target nucleic acid but does not promote a nucleic acid extension reaction unless UV light is irradiated, a reaction field area arranged with a plurality of said nucleic acid probes that hybridize with different target nucleic acids, and a nucleic acid analysis device having a channel that is installed with a plurality of said reaction field areas, and that is capable of retaining a reagent to be utilized in the nucleic acid reaction; an excited optical system for irradiating the evanescent light to the desired reaction field area and detecting fluorescence generated; a reaction control optical system for irradiating the UV light to decompose the photodegradable substance to the desired reaction field area; and a stage drive mechanism for moving relatively the nucleic acid analysis device to the excited optical system and the reaction control optical system, so as to irradiate evanescent light and UV light to the desired reaction field area. - View Dependent Claims (10, 11, 12, 13, 14, 15, 16)
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17. A single molecule real time sequencing method comprising:
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preparing a nucleic acid probe having a photodegradable substance that inhibits a nucleic acid extension reaction and a channel installed with a plurality of reaction field areas arranged with a plurality of said nucleic acid probes; supplying a target nucleic acid and a reagent to be utilized in the nucleic acid extension reaction into the channel; irradiating UV light to decompose the photodegradable substance to the desired reaction field area; irradiating evanescent light to the desired reaction field area, and detecting fluorescence generated; moving relatively the channel to the reaction control optical system for irradiating the UV light, and the excited optical system generating the evanescent light; irradiating UV light to decompose the photodegradable substance to the different reaction field areas; and irradiating evanescent light to the different reaction field area, and to detect fluorescence generated. - View Dependent Claims (18, 19, 20, 21, 22, 23)
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Specification