Assembly for label-free detection of hybridized nucleic targets
First Claim
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1. An assembly for performing an assay, the assembly comprises:
- a) an article with a solid support upon which is immobilized a number of first probes, each of said first probes having a second probe attached thereto, forming a duplex, such that each first probe includes a first nucleic acid sequence to which an emitter element is attached, and each second probe includes a second nucleic acid sequence, which is a complementary sequence to said first nucleic acid sequence and containing at least one artificial mismatch site, and to which a quenching element is attached; and
b) a reagent solution comprising an unlabeled sample nucleic acid target, which has a nucleic sequence that is perfectly complementary to said first-probe nucleic acid sequence and is adapted to displace said second probe from said first probe in competitive hybridization, whereby said emitter element increases in signal intensity.
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Abstract
An assembly to practice a method for the detection and analysis of genetic polymorphisms using arrays that do not require labeling of a target nucleic acid sequence. Hybridization of a perfectly complementary nucleic acid target sequence to an oligonucleotide probe sequence results in a displacement and complete removal of a hybridized probe sequence from the same oligonucleotide probe sequence by means of a thermo-kinetic reaction. The removal of the hybridized probe sequence, having a quencher element, increases the intensity of emission by an emitter element on the oligonucleotide probe sequence.
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10 Claims
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1. An assembly for performing an assay, the assembly comprises:
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a) an article with a solid support upon which is immobilized a number of first probes, each of said first probes having a second probe attached thereto, forming a duplex, such that each first probe includes a first nucleic acid sequence to which an emitter element is attached, and each second probe includes a second nucleic acid sequence, which is a complementary sequence to said first nucleic acid sequence and containing at least one artificial mismatch site, and to which a quenching element is attached; and
b) a reagent solution comprising an unlabeled sample nucleic acid target, which has a nucleic sequence that is perfectly complementary to said first-probe nucleic acid sequence and is adapted to displace said second probe from said first probe in competitive hybridization, whereby said emitter element increases in signal intensity. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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10. An assembly for performing an assay, the assembly comprises:
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a) an article with a solid support upon which is immobilized a number of first probes, each first probe having a first nucleic acid sequence to which an emitter element is attached; and
b) a reagent solution comprising
1) a second probe and
2) an unlabeled sample nucleic acid target, both of which are adapted to hybridize with said first probe under competitive hybridization conditions, wherein said second probe has a second nucleic acid sequence that is a complementary sequence to said first-probe nucleic acid sequence and containing at least one artificial mismatch site, and to which a quenching element is attached, and said nucleic acid target has a nucleic sequence that is perfectly complementary to said first-probe nucleic acid sequence.
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Specification