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Title: 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:
 [1];  [2]
  1. Santa Fe, NM
  2. Los Alamos, NM
Issue Date:
Research Org.:
Los Alamos National Lab. (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 = {2006},
month = {12}
}

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Works referenced in this record:

A ligase-mediated gene detection technique
journal, August 1988


Universal DNA array detection of small insertions and deletions in BRCA1 and BRCA2
journal, May 2000


Detection of specific alleles by using allele-specific primer extension followed by capture on solid support
journal, August 1992


Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide‚ÄČArrays
journal, June 1997