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Title: Dual phase multiplex polymerase chain reaction

Abstract

Highly specific and sensitive methods were developed for multiplex amplification of nucleic acids on supports such as microarrays. Based on a specific primer design, methods include five types of amplification that proceed in a reaction chamber simultaneously. These relate to four types of multiplex amplification of a target DNA on a solid support, directed by forward and reverse complex primers immobilized to the support and a fifth type--pseudo-monoplex polymerase chain reaction (PCR) of multiple targets in solution, directed by a single pair of unbound universal primers. The addition of the universal primers in the reaction mixture increases the yield over the traditional "bridge" amplification on a solid support by approximately ten times. Methods that provide multitarget amplification and detection of as little as 0.45-4.5.times.10.sup.-12 g (equivalent to 10.sup.2-10.sup.3 genomes) of a bacterial genomic DNA are disclosed.

Inventors:
 [1];  [2]
  1. Charlottesville, VA
  2. Darien, IL
Issue Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
985411
Patent Number(s):
7432055
Application Number:
10/794,381
Assignee:
UChicago Argonne LLC (Chicago, IL)
Patent Classifications (CPCs):
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
DOE Contract Number:  
W-31-109-ENG-38
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Pemov, Alexander, and Bavykin, Sergei. Dual phase multiplex polymerase chain reaction. United States: N. p., 2008. Web.
Pemov, Alexander, & Bavykin, Sergei. Dual phase multiplex polymerase chain reaction. United States.
Pemov, Alexander, and Bavykin, Sergei. Tue . "Dual phase multiplex polymerase chain reaction". United States. https://www.osti.gov/servlets/purl/985411.
@article{osti_985411,
title = {Dual phase multiplex polymerase chain reaction},
author = {Pemov, Alexander and Bavykin, Sergei},
abstractNote = {Highly specific and sensitive methods were developed for multiplex amplification of nucleic acids on supports such as microarrays. Based on a specific primer design, methods include five types of amplification that proceed in a reaction chamber simultaneously. These relate to four types of multiplex amplification of a target DNA on a solid support, directed by forward and reverse complex primers immobilized to the support and a fifth type--pseudo-monoplex polymerase chain reaction (PCR) of multiple targets in solution, directed by a single pair of unbound universal primers. The addition of the universal primers in the reaction mixture increases the yield over the traditional "bridge" amplification on a solid support by approximately ten times. Methods that provide multitarget amplification and detection of as little as 0.45-4.5.times.10.sup.-12 g (equivalent to 10.sup.2-10.sup.3 genomes) of a bacterial genomic DNA are disclosed.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2008},
month = {10}
}

Works referenced in this record:

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Manual Manufacturing of Oligonucleotide, DNA, and Protein Microchips
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An oligonucleotide hybridization approach to DNA sequencing
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PCR Amplification on a Microarray of Gel-Immobilized Oligonucleotides: Detection of Bacterial Toxin- and Drug-Resistant Genes and Their Mutations
journal, October 2000


DNA analysis and diagnostics on oligonucleotide microchips.
journal, May 1996


Integration of Multiple PCR Amplifications and DNA Mutation Analyses by Using Oligonucleotide Microchip
journal, May 2001


Synthesis and properties of oligodeoxyribonucleotide—polyethylene glycol conjugates
journal, January 1994