Title:
単一核酸分子の塩基配列決定法
Document Type and Number:
Japanese Patent JP4721606
Kind Code:
B2
Abstract:
The present invention relates to a method for determining a nucleotide sequence of a nucleic acid by single dye molecule detection, the method comprising: (1) a step of immobilizing a nucleic acid molecule, or a primer which has a sequence complementary to a part of the sequence of the nucleic acid molecule, onto the surface of a solid; (2) a step of annealing the primer or the nucleic acid molecule to the nucleic acid molecule or the primer, respectively; (3) a step of providing a solution which contains DNA polymerase and one type of dye-labeled dNTP, or RNA polymerase and one type of dye-labeled NTP, to the immobilized nucleic acid molecule, and allowing the nucleotide to react with the 3' end of the primer, whereby a nucleotide, which forms a base-pair with a base opposed to the reaction site, is bound to the primer by action of the polymerase; (4) a step of detecting the presence of a bound, dye-labeled dNTP or NTP; (5) a step of disrupting the dye molecule of the bound, dye-labeled dNTP or NTP; (6) a step of repeating the steps (3) to (5) while changing the type of dye-labeled dNTP or NTP in turn, to sequentially bind dNTPs or NTPs which base-pair with the nucleotides of the nucleic acid molecule; and (7) a step of determining the nucleotide sequence of the nucleic acid molecule based on the types of the sequentially bound dNTPs or NTPs.
Inventors:
Yoshihiro Takeda
Fumitaka Manafune
Yasushi Kondo
Fumitaka Manafune
Yasushi Kondo
Application Number:
JP2001570834A
Publication Date:
July 13, 2011
Filing Date:
March 30, 2001
Export Citation:
Assignee:
TOYOTA JIDOSHA KABUSHIKI KAISHA
Genesis Research Institute, Inc.
Genesis Research Institute, Inc.
International Classes:
C12N15/09; C12Q1/68; G01N33/50; G01N33/58; C12M1/00
Domestic Patent References:
| JP7507681A |
Foreign References:
| US5846727A | 1998-12-08 |
Attorney, Agent or Firm:
Yusuke Hiraki
Ishii Teiji
Ishii Teiji