Nucleic acid sequence detection using multiplexed oligonucleotide PCR
Abstract
Methods for rapidly detecting single or multiple sequence alleles in a sample nucleic acid are described. Provided are all of the oligonucleotide pairs capable of annealing specifically to a target allele and discriminating among possible sequences thereof, and ligating to each other to form an oligonucleotide complex when a particular sequence feature is present (or, alternatively, absent) in the sample nucleic acid. The design of each oligonucleotide pair permits the subsequent high-level PCR amplification of a specific amplicon when the oligonucleotide complex is formed, but not when the oligonucleotide complex is not formed. The presence or absence of the specific amplicon is used to detect the allele. Detection of the specific amplicon may be achieved using a variety of methods well known in the art, including without limitation, oligonucleotide capture onto DNA chips or microarrays, oligonucleotide capture onto beads or microspheres, electrophoresis, and mass spectrometry. Various labels and address-capture tags may be employed in the amplicon detection step of multiplexed assays, as further described herein.
- Inventors:
-
- Santa Fe, NM
- Los Alamos, NM
- Issue Date:
- Research Org.:
- Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 903353
- Patent Number(s):
- 7153656
- Application Number:
- 10/336,266
- Assignee:
- Los Alamos National Security, LLC (Los Alamos, NM)
- Patent Classifications (CPCs):
-
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
- DOE Contract Number:
- W-7405-ENG-36
- Resource Type:
- Patent
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Nolan, John P, and White, P Scott. Nucleic acid sequence detection using multiplexed oligonucleotide PCR. United States: N. p., 2006.
Web.
Nolan, John P, & White, P Scott. Nucleic acid sequence detection using multiplexed oligonucleotide PCR. United States.
Nolan, John P, and White, P Scott. Tue .
"Nucleic acid sequence detection using multiplexed oligonucleotide PCR". United States. https://www.osti.gov/servlets/purl/903353.
@article{osti_903353,
title = {Nucleic acid sequence detection using multiplexed oligonucleotide PCR},
author = {Nolan, John P and White, P Scott},
abstractNote = {Methods for rapidly detecting single or multiple sequence alleles in a sample nucleic acid are described. Provided are all of the oligonucleotide pairs capable of annealing specifically to a target allele and discriminating among possible sequences thereof, and ligating to each other to form an oligonucleotide complex when a particular sequence feature is present (or, alternatively, absent) in the sample nucleic acid. The design of each oligonucleotide pair permits the subsequent high-level PCR amplification of a specific amplicon when the oligonucleotide complex is formed, but not when the oligonucleotide complex is not formed. The presence or absence of the specific amplicon is used to detect the allele. Detection of the specific amplicon may be achieved using a variety of methods well known in the art, including without limitation, oligonucleotide capture onto DNA chips or microarrays, oligonucleotide capture onto beads or microspheres, electrophoresis, and mass spectrometry. Various labels and address-capture tags may be employed in the amplicon detection step of multiplexed assays, as further described herein.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue Dec 26 00:00:00 EST 2006},
month = {Tue Dec 26 00:00:00 EST 2006}
}
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