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Apparatus for determining base sequence of nucleic acid

  • US 6,242,193 B1
  • Filed: 05/09/2000
  • Issued: 06/05/2001
  • Est. Priority Date: 07/30/1999
  • Status: Expired due to Fees
First Claim
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1. An apparatus for determining a base sequence of a template DNA comprising:

  • a cell in which a complementary strand extension reaction using a polymerase is performed for extending a hybridized primer hybridized with a template DNA or extending an extended primer produced by said complementary strand extension reaction, by incorporating a single chemically modified nucleotide of four kinds of chemically modified nucleotides to 3′

    -terminus of said hybridized primer or said extended primer, in the presence of said four kinds of chemically modified nucleotides, wherein said single chemically modified nucleotide is complementary with a base sequence of said template DNA, and each of said four kinds of chemically modified nucleotides has a chemical modification for preventing a continuous progress of said complementary strand extension reaction, after incorporating said single chemically modified nucleotide to 3′

    -terminus of said hybridized primer or said extended primer;

    a laser which excites at least one fluorophore label included in said chemical modification of said chemically modified nucleotide incorporated to 3′

    -terminus of said hybridized primer or said extended primer;

    a lens which collects fluorescence emitted from said fluorophore label(s);

    filters, each of which selects a wavelength of said fluorescence;

    a photodetector which detects said fluorescence selected by said filters;

    an analyzer which determines a kind of a base of said chemically modified nucleotide incorporated to 3′

    -terminus of said hybridized primer or said extended primer, based on an output of said photodetector; and

    a light source which emits a light for releasing said chemical modification from said chemically modified nucleotide incorporated to 3′

    -terminus of said hybridized primer or said extended primer, in order to bring about a state at which said complementary strand extension reaction for extending said hybridized primer or said extended primer can proceed;

    wherein, by repeating said complementary strand extension reaction for extending said hybridized primer or said extended primer and releasing said chemical modification from said chemically modified nucleotide incorporated to 3′

    -terminus of said hybridized primer or said extended primer, said base sequence of said template DNA is determined one base by one base sequentially form the kinds of bases of said chemically modified nucleotides incorporated to said extended primer by said complementary strand extension reaction.

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