Method and kit for analyzing target nucleic acid fragment
First Claim
1. A kit for analyzing a target nucleic acid fragment, comprising a reagent for polymerase extension reaction including at least one primer that is free in solution and complementary to a target nucleic acid fragment to be analyzed, at least one deoxyribonucleotide triphosphate and at least one polymerase, and an electrochemically active threading intercalator which intercalates to a double-stranded nucleic acid and allows a shift of the electrochemical response.
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Abstract
A method of analyzing a target nucleic acid fragment is provided that is capable of being performed simply and promptly, by anyone, using a compact device without requirement of specific technique or complex operation. The method comprises timely detection of a double-stranded nucleic acid fragment formed during the polymerase extension reaction due to a specific base sequence of the target nucleic acid fragment as increase of an electrochemical response under the existence of an electrochemically active intercalator; and the kit utilizes the method.
9 Citations
12 Claims
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1. A kit for analyzing a target nucleic acid fragment, comprising a reagent for polymerase extension reaction including at least one primer that is free in solution and complementary to a target nucleic acid fragment to be analyzed, at least one deoxyribonucleotide triphosphate and at least one polymerase, and an electrochemically active threading intercalator which intercalates to a double-stranded nucleic acid and allows a shift of the electrochemical response.
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2. A method for analyzing a target nucleic acid fragment of which at least a part of base sequence has been known, comprising:
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contacting the target nucleic acid fragment with a primer that is free in solution and complementary to the target nucleic acid fragment to form into a double-stranded nucleic acid fragment; extending the primer from a terminal by polymerase extension reaction with deoxyribonucleotide triphosphate in the presence of a polymerase utilizing the target nucleic acid fragment as a template; contacting an electrochemically active intercalator to intercalate to the double-stranded nucleic acid fragment extended by the polymerase extension reaction where the electrochemically active intercalator is a threading intercalator having redox activity; and detecting the double-stranded nucleic acid fragment by measuring shift of electrochemical response. - View Dependent Claims (3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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Specification