Method of identifying nucleic acid
OwnerJoji Oshima, Ken Nemotoinventors
Life of this patent
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Abstract
Test nucleic acids are identified by the steps of: synthesizing multiple nucleic acids that comprise nucleotide sequences complementary to different regions of a test nucleic acid; and comparing the dissociation curve of a mixture of the synthesized nucleic acids. The dissociation curve of a mixture of nucleic acids shows a waveform pattern that is unique to the test nucleic acid. Many types of nucleic acids can be efficiently identified using simple reactions. The multiple types of nucleic acids that are necessary for analysis can be easily synthesized by using a primer complex that anneals to multiple regions of the test nucleic acid.
Claims58
A method for identifying a nucleic acid, wherein the method comprises the steps of:
- (1) synthesizing a nucleic acid comprising a nucleotide sequence complementary to multiple regions of a test nucleic acid;
- (2) obtaining dissociation curves for a mixture of the nucleic acid synthesized in step (1);
- and
- (3) comparing the waveform patterns of the dissociation curves and identifying nucleic acids comprising the same waveform pattern to have the same nucleotide sequence.
A method for producing dissociation curve waveform patterns as a reference for identifying a nucleic acid, wherein the method comprises the steps of:
- (1) synthesizing nucleic acids, which comprise nucleotide sequences complementary to multiple regions of a standard nucleic acid sample, as test nucleic acids;
- and
- (2) obtaining dissociation curves for the mixture of nucleic acids synthesized in step (1).
A primer complex for waveform production, which comprises a mixture of one or more types of primers whose nucleotide sequences are complementary to multiple regions in a test nucleic acid.
A method for producing a primer complex for waveform production, wherein the complex comprises:
- a specific primer, which comprises a nucleotide sequence complementary to a target region of a template nucleic acid;
- and
- at least one type of ambiguous primer, which comprise the following specific region and ambiguous region:
- a specific region, which comprises the 3′ end of a primer and consists of a nucleotide sequence complementary to the target region;
- and
- an ambiguous region, which is positioned to the 5′ side of the specific region, and comprises a nucleotide sequence wherein the nucleotide comprised in the nucleotide sequence complementary to the target region is substituted with a nucleotide other than the nucleotide;
- and
- wherein the method comprises the steps of:
- a) synthesizing the specific region;
- and
- b) synthesizing the ambiguous region by binding a nucleotide of a nucleotide sequence complementary to the target region, to a mixture of random nucleotides other than said nucleotide, selected from adenine, cytosine, guanine, and thymine.
A kit for identifying a nucleic acid, wherein the kit comprises the components of:
- (1) a primer complex for waveform production, that comprises a mixture of one or more types of primers comprising nucleotide sequences complementary to multiple regions of a test nucleic acid;
- (2) a DNA polymerase that catalyzes template-specific synthesis of a complementary strand;
- and
- (3) a substrate for complementary strand synthesis.
Dependencies are inferred from the claim text.