Compositions and methods for free-solution conjugate nucleic acid analysis
First Claim
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1. A method for the detection of single nucleotide polymorphisms comprising:
- a) providing one or more DNA samples to be tested,b) providing eight or more DNA-polyamide bioconjugates specific to one or more single nucleotide polymorphisms, wherein said eight or more DNA-polyamide bioconjugates each comprise a primer sequence linked to one of eight or more different polyamide drag-tags that differ in size, wherein said eight or more different polyamide drag-tags comprise methoxyethylglycine (NMEG) monomers, and wherein each of said eight or more different polyamide drag-tags has a size that differs by no more than 4 NMEG monomers from the size of the other drag-tag(s) closest in size;
c) contacting said DNA sample with said DNA-polyamide bioconjugates,d) performing a single nucleotide polymorphism assay on said DNA sample, wherein said assay is conducted under conditions that generate reaction products that are individually resolvable in a free-solution electrophoresis instrument,e) applying said DNA sample to a free-solution electrophoretic instrument under conditions such that said eight or more DNA-polyaminde bioconjugates are separated from each other in said free-solution electrophoretic instrument, andf) detecting the presence or absence of a single nucleotide polymorphism in said DNA sample.
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Abstract
The present invention provides compositions and methods for performing free-solution conjugate analysis of nucleic acid molecules. For example, the present invention provides multiplexed single-base extension assays for genotyping. In particular, the present invention provides a series of disperse polyamide “drag tags” for use in achieving high-resolution separation of nucleic acid reaction products.
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2 Claims
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1. A method for the detection of single nucleotide polymorphisms comprising:
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a) providing one or more DNA samples to be tested, b) providing eight or more DNA-polyamide bioconjugates specific to one or more single nucleotide polymorphisms, wherein said eight or more DNA-polyamide bioconjugates each comprise a primer sequence linked to one of eight or more different polyamide drag-tags that differ in size, wherein said eight or more different polyamide drag-tags comprise methoxyethylglycine (NMEG) monomers, and wherein each of said eight or more different polyamide drag-tags has a size that differs by no more than 4 NMEG monomers from the size of the other drag-tag(s) closest in size; c) contacting said DNA sample with said DNA-polyamide bioconjugates, d) performing a single nucleotide polymorphism assay on said DNA sample, wherein said assay is conducted under conditions that generate reaction products that are individually resolvable in a free-solution electrophoresis instrument, e) applying said DNA sample to a free-solution electrophoretic instrument under conditions such that said eight or more DNA-polyaminde bioconjugates are separated from each other in said free-solution electrophoretic instrument, and f) detecting the presence or absence of a single nucleotide polymorphism in said DNA sample. - View Dependent Claims (2)
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Current AssigneeNorthwestern University
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Original AssigneeNorthwestern University
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InventorsMeagher, Robert J., Barron, Annelise E., Vreeland, Wyatt N
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Primary Examiner(s)Kim, Young J
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Assistant Examiner(s)BERTAGNA, ANGELA MARIE
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Application NumberUS11/960,185Publication NumberTime in Patent Office1,518 DaysField of SearchNoneUS Class Current435/6.11CPC Class CodesC12Q 1/6827 for detection of mutation o...C12Q 2533/101 Primer extensionC12Q 2565/125 Electrophoretic separationC12Q 2565/627 being a mass spectrometerY10T 436/143333 Saccharide [e.g., DNA, etc.]
