COMPOSITIONS AND METHODS FOR FREE-SOLUTION CONJUGATE NUCLEIC ACID ANALYSIS

• US 20080227211A1
• Filed: 12/19/2007
• Published: 09/18/2008
• Est. Priority Date: 12/19/2006
• Status: Active Grant

• Current Assignee: Northwestern University
• Original Assignee: Northwestern University
• Inventors: Meagher, Robert J., Vreeland, Wyatt N., Barron, Annelise E.
• Granted Patent: US 8,114,599 B2
• Time in Patent Office: Days
• US Class Current: 436/94
• CPC Class Codes: C12Q 1/6827 for detection of mutation or polymorphism
• CPC Class Codes: C12Q 2533/101 Primer extension
• CPC Class Codes: C12Q 2565/125 Electrophoretic separation
• CPC Class Codes: C12Q 2565/627 being a mass spectrometer
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

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.

Claims

1. A composition comprising a set of four or more monodisperse uncharged polyamides conjugated to four or more DNA molecules such that the electrophoretic mobility of the DNA-polyamide bioconjugate is modified compared to an unconjugated DNA molecule.

3. The composition of claim 1, wherein said bioconjugate comprises uncharged polyamides conjugated to both ends of said DNA molecule.

2. The composition of claim 2, wherein said bioconjugate comprises an uncharged polyamide conjugated to one end of said DNA molecule.

4. A method for detection of single nucleotide polymorphisms comprising:

a) providing one or more DNA samples to be tested,

b) providing four or more DNA-polyamide bioconjugates specific to one or more single nucleotide polymorphisms,

c) contacting said DNA sample with said DNA-polyamide bioconjugates,

d) performing a single nucleotide polymorphism assay under conditions such that a single nucleotide polymorphism in said DNA sample is detected if present,

e) applying the assayed sample to a free solution electrophoretic instrument, and

f) detecting the presence or absence of a single nucleotide polymorphism in said DNA sample.

5. The method of claim 4, further comprising the determination of a disease state in said subject DNA sample based on the detection of the presence of a single nucleotide polymorphism.

6. The method of claim 4, further comprising the diagnosis of a disease state in said subject based on the detection of the presence of a disease state single nucleotide polymorphism in said subject DNA sample.