Method for nucleic acid analysis
First Claim
1. A method for nucleic acid analysis to analyze one or more noncomplementary sites in a double-stranded nucleic acid sample, comprising the steps of:
- obtaining nucleic acid fragments by cleaving the one or more noncomplementary sites with a single strand-specific endonuclease;
hybridizing at least one of the obtained nucleic acid fragments to a probe having a nucleotide sequence that is partially or totally identical to either one strand of the double-stranded nucleic acid sample;
performing an extension reaction from the nucleic acid fragment hybridized to the probe; and
optically detecting pyrophosphate generated by the extension reaction.
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Abstract
A simple and highly accurate method for detecting the presence or absence of gene mutation and methylated cytosine in CpG dinucleotide that are contained in a target sequence derived from an analysis sample is provided. Features of the method for nucleic acid analysis include cleaving one or more noncomplementary sites in a double-stranded nucleic acid sample by a single strand-specific endonuclease, hybridizing at least one of the nucleic acid fragments obtained to a probe containing a nucleotide sequence that is partially or totally identical to either one strand of the double-stranded nucleic acid sample, allowing an extension reaction to proceed from the nucleic acid fragment hybridized to the probe, and optically detecting pyrophosphate generated by the extension reaction, thereby judging the presence or absence of at least a noncomplementary site in the double-stranded nucleic acid sample.
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Citations
12 Claims
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1. A method for nucleic acid analysis to analyze one or more noncomplementary sites in a double-stranded nucleic acid sample, comprising the steps of:
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obtaining nucleic acid fragments by cleaving the one or more noncomplementary sites with a single strand-specific endonuclease;
hybridizing at least one of the obtained nucleic acid fragments to a probe having a nucleotide sequence that is partially or totally identical to either one strand of the double-stranded nucleic acid sample;
performing an extension reaction from the nucleic acid fragment hybridized to the probe; and
optically detecting pyrophosphate generated by the extension reaction. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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Specification