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US 2007/0190543 A1Published application
Coded Molecules for Detecting Target Analytes
AssigneeApplera Corporation
InventorKenneth Livak
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Abstract
The present disclosure relates to methods of detecting target analytes based on single molecule detection of coded molecules.
Claims86
A method of detecting a target analyte, comprising:
- (a) translocating a coded molecule through a nanopore, wherein the coded molecule comprises (i) an ordered plurality of code regions formed on a single-stranded nucleobase polymer scaffold, wherein one or more of the code regions is non-single-stranded and each code region has a detectable property such that detecting the ordered code regions generates a defined signal pattern, and (ii) a moiety capable of binding to a target analyte, and
- wherein the nanopore is dimensioned for passage of the non-single-stranded region;
- (b) sensing the detectable property of each code region as the coded molecule translocates through the nanopore to generate the defined signal pattern;
- and
- (c) relating the signal pattern to presence of the moiety.
2 more elements
A method of detecting a target analyte, comprising:
- (a) translocating a coded molecule of a population of coded molecules through a nanopore, wherein the population of coded molecules comprises at least a first and second subpopulation, the coded molecule of the population comprising:
- (i) an ordered plurality of code regions formed on a single-stranded nucleobase polymer scaffold, wherein one or more of the code regions is non-single-stranded and each code region has a detectable property such that detecting a combination of code regions generates a defined signal pattern distinguishable between the first and second subpopulations;
- and
- (ii) a first moiety on the first subpopulation, wherein the first moiety is capable of binding to a first target analyte, and a second moiety on the second subpopulation, wherein the second moiety is capable of binding to a second target analyte, and
- wherein the nanopore is of a sufficient dimension for transit of the non-single-stranded region;
- (b) sensing the detectable property of each code region as the coded molecule translocates through the nanopore to generate the defined signal pattern;
- and
- (c) relating the signal pattern to the presence of the moiety of the first or second subpopulation.
5 more elements
A method of detecting a target analyte, comprising:
- (a) translocating a coded molecule of a population of coded molecules through a nanopore, wherein the population of coded molecules comprises a plurality of subpopulations, the coded molecule of each subpopulation comprising:
- (i) an ordered plurality of code regions formed on a single-stranded nucleobase polymer scaffold, wherein one or more of the code regions is non-single-stranded and each code region comprises a detectable property such that the ordered code regions generates a defined signal pattern characteristic for each subpopulation;
- and
- (ii) a moiety capable of binding to a target analyte, wherein the moiety of each subpopulation binds to a different target analyte, and
- wherein the nanopore is dimensioned for transit of the non-single-stranded region;
- (b) sensing the detectable property of each code region translocating through the nanopore to generate the signal pattern;
- and
- (c) relating the generated signal pattern to the moiety of the subpopulation.
5 more elements
A method of forming a coded molecule, comprising:
- (a) contacting a microcapsule with a first nucleobase oligomer, wherein the microcapsule comprises a single-stranded nucleobase scaffold;
- and
- (b) hybridizing the first nucleobase oligomer to the scaffold, wherein the first nucleobase oligomer hybridizes to a first defined sequence on the scaffold to form a first non-single-stranded code region.
A method of forming a coded molecule, comprising
- (a) contacting a population of microcapsules with a first nucleobase oligomer, wherein the microcapsules comprise a single-stranded nucleobase scaffold and the first nucleobase oligomer hybridizes to a first defined sequence on the scaffold to form a first non-single-stranded code region;
- (b) generating from the population of microcapsules at least a first and second subpopulation of microcapsules;
- and
- (c) contacting the first subpopulation of microcapsules with a second nucleobase oligomer and the third subpopulation with a third nucleobase oligomer, wherein the second nucleobase oligomer hybridizes to a second defined sequence and the third nucleobase oligomer hybridizes to a third defined sequence on the scaffold to form a second non-single-stranded code region in each subpopulation.
1 more element
A method of forming a coded molecule, comprising
- (a) contacting a population of microcapsules with a first nucleobase oligomer, wherein the microcapsules comprise a single-stranded nucleobase scaffold and the first nucleobase oligomer hybridizes to a first defined sequence on the scaffold to form a first non-single-stranded code region;
- (b) generating from the population of microcapsules a plurality of microcapsule subpopulations;
- (c) contacting each subpopulation with a second nucleobase oligomer that hybridizes to a second defined sequence on the scaffold to form a second non-single-stranded code region, wherein the second nucleobase oligomer of each subpopulation comprises a different nucleobase polymer.
Dependencies are inferred from the claim text.