# US 2013/0157264 A1 · Nucleic acid analysis device, nucleic acid analysis apparatus, and nucleic acid analysis method > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2013/0157264 A1 - Document ID: US20130157264A1 - Status: Published application - Kind: A1 - Application number: 13/818,704 - Filed: 2011-07-19 - Published: 2013-06-20 - Earliest claimed: 2010-09-10 - Inventors: Takayuki Obara; Kazumichi Imai; Toshiro Saito; Satoshi Takahashi - Web page: https://patentgrep.com/patent/US20130157264A1 ## CPC - G01N 21/6486: {Measuring fluorescence of biological material, e.g. DNA, RNA, cells (G01N21/6428 takes precedence)} - C12Q 1/6837: using probe arrays or probe chips (C12Q1/6874 takes precedence) - G01N 21/6428: {Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53)} - G01N 21/6452: {Individual samples arranged in a regular 2D-array, e.g. multiwell plates} - G01N 21/648: {using evanescent coupling or surface plasmon coupling for the excitation of fluorescence} - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES ## Abstract In a nucleic acid analysis device which detects a fluorescent dye on a nucleic acid sample immobilized on a surface of a substrate by exciting the fluorescent dye with an evanescent wave, the detection of a fluorescence signal with a high SN ratio is realized even for a long nucleic acid sample. The nucleic acid analysis device according to the invention is a nucleic acid analysis device in which a plurality of regions for immobilizing a nucleic acid sample are provided on a surface of a support base and a single molecule of a nucleic acid sample is immobilized on at least one of the regions, and which performs sequence determination by performing an extension reaction of the immobilized nucleic acid sample, wherein the immobilization of the single molecule of the nucleic acid sample on the support base is performed at two or more points. ## Claims ### Claim 1 A nucleic acid analysis device, in which a plurality of regions for immobilizing a nucleic acid sample are provided on a surface of a support base and a single molecule of a nucleic acid sample is immobilized on at least one of, the regions, and which performs sequence determination by performing an extension reaction of the immobilized nucleic acid sample, wherein the immobilization of the single molecule of the nucleic acid sample on the support base is performed at two or more points. ### Claim 2 (depends on 1) The nucleic acid analysis device according to claim 1, wherein the regions are subjected to a surface treatment so that the nucleic acid sample can be immobilized thereon. ### Claim 3 (depends on 2) The nucleic acid analysis device according to claim 2, wherein two or more different surface treatments are performed for at least one of the regions. ### Claim 4 (depends on 2) The nucleic acid analysis device according to claim 2, wherein the surface treatment varies between at least some of the regions. ### Claim 5 (depends on 1) The nucleic acid analysis device according to claim 1, wherein a metal structure is disposed in the regions. ### Claim 6 (depends on 5) The nucleic acid analysis device according to claim 5, wherein a microparticle is immobilized on the metal structure. ### Claim 7 (depends on 6) The nucleic acid analysis device according to claim 6, wherein the metal structure has a circular planar shape and has a diameter smaller than that of the microparticle. ### Claim 8 (depends on 5) The nucleic acid analysis device according to claim 5, wherein the metal structure is made of any of gold, silver, aluminum, chromium, titanium, tungsten, platinum, and nickel. ### Claim 9 (depends on 1) The nucleic acid analysis device according to claim 1, wherein the regions are arranged in a lattice pattern. ### Claim 10 (depends on 1) The nucleic acid analysis device according to claim 1, wherein at least one end of the nucleic acid sample is immobilized on the support base at the 3′ end. ### Claim 11 (depends on 2) The nucleic acid analysis device according to claim 2, wherein the surface treatment is a nucleic acid molecule immobilized on the surface of the support base. ### Claim 12 A nucleic acid analysis method in which sequence determination is performed by immobilizing a single molecule of a nucleic acid sample in at least one of a plurality of regions for immobilizing a nucleic acid sample on a surface of a support base and performing an extension reaction of the immobilized nucleic acid sample, wherein the immobilization of the single molecule of the nucleic acid sample to the support base is performed at two or more points. ### Claim 13 A nucleic acid analysis method comprising: a step of performing a nucleic acid synthesis reaction by bringing a nucleic acid synthetase and bases with a fluorescent dye into contact with a nucleic acid sample; and a step of detecting fluorescence generated by exciting a label by local illumination, wherein the method includes: a step of immobilizing a single molecule on a surface of a support base at a first point of a nucleic acid sample; and a step of immobilizing the molecule on the surface of the support base at a second point of the nucleic acid sample. ### Claim 14 (depends on 13) The nucleic acid analysis method according to claim 13, wherein a step including the step of detecting fluorescence generated by excitation through local illumination is performed in a state where the bases with a fluorescent dye are present around the nucleic acid sample. ### Claim 15 (depends on 13) The nucleic acid analysis method according to claim 13, wherein a phosphor is bound to the nucleic acid synthetase. ### Claim 16 (depends on 13) The nucleic acid analysis method according to claim 13, wherein the immobilization of the nucleic acid sample on the support base is achieved through both ends of one strand of the nucleic acid sample. ### Claim 17 (depends on 13) The nucleic acid analysis method according to claim 13, wherein the immobilization of the nucleic acid sample on the support base is achieved through a covalent bond. ### Claim 18 (depends on 13) The nucleic acid analysis method according to claim 13, wherein the immobilization of the nucleic acid sample on the support base is achieved through a protein. ### Claim 19 (depends on 13) The nucleic acid analysis method according to claim 13, wherein the immobilization of the nucleic acid sample on the support base is performed at three or more points. ### Claim 20 (depends on 13) The nucleic acid analysis method according to claim 13, wherein the method includes a step of introducing a modification into both ends of the nucleic acid sample. ### Claim 21 A nucleic acid analysis apparatus which determines a base sequence of a nucleic acid sample and comprises: a nucleic acid analysis device in which a plurality of regions for immobilizing a nucleic acid sample are provided on a surface of a support base and a single molecule of a nucleic acid sample is immobilized on at least one of the regions; a section which supplies at least a nucleic acid sample to the nucleic acid analysis device; a section which irradiates the nucleic acid analysis device with light; and a detection section which detects fluorescence incorporated by a nucleic acid extension reaction, wherein the immobilization of the single molecule of the nucleic acid sample on the support base is performed at two or more points. ### Claim 22 (depends on 21) The nucleic acid analysis apparatus according to claim 21, wherein the regions are arranged in a lattice pattern.