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Title: Flow cytometric detection method for DNA samples

Abstract

Disclosed herein are two methods for rapid multiplex analysis to determine the presence and identity of target DNA sequences within a DNA sample. Both methods use reporting DNA sequences, e.g., modified conventional Taqman.RTM. probes, to combine multiplex PCR amplification with microsphere-based hybridization using flow cytometry means of detection. Real-time PCR detection can also be incorporated. The first method uses a cyanine dye, such as, Cy3.TM., as the reporter linked to the 5' end of a reporting DNA sequence. The second method positions a reporter dye, e.g., FAM, on the 3' end of the reporting DNA sequence and a quencher dye, e.g., TAMRA, on the 5' end.

Inventors:
 [1];  [1];  [1]
  1. Livermore, CA
Issue Date:
Research Org.:
Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
908290
Patent Number(s):
7083951
Application Number:
W-7405-ENG-48
Assignee:
The Regents of the University of California (Oakland, CA)
Patent Classifications (CPCs):
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
DOE Contract Number:  
W-7405-ENG-48
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Nasarabadi, Shanavaz, Langlois, Richard G, and Venkateswaran, Kodumudi S. Flow cytometric detection method for DNA samples. United States: N. p., 2006. Web.
Nasarabadi, Shanavaz, Langlois, Richard G, & Venkateswaran, Kodumudi S. Flow cytometric detection method for DNA samples. United States.
Nasarabadi, Shanavaz, Langlois, Richard G, and Venkateswaran, Kodumudi S. Tue . "Flow cytometric detection method for DNA samples". United States. https://www.osti.gov/servlets/purl/908290.
@article{osti_908290,
title = {Flow cytometric detection method for DNA samples},
author = {Nasarabadi, Shanavaz and Langlois, Richard G and Venkateswaran, Kodumudi S},
abstractNote = {Disclosed herein are two methods for rapid multiplex analysis to determine the presence and identity of target DNA sequences within a DNA sample. Both methods use reporting DNA sequences, e.g., modified conventional Taqman.RTM. probes, to combine multiplex PCR amplification with microsphere-based hybridization using flow cytometry means of detection. Real-time PCR detection can also be incorporated. The first method uses a cyanine dye, such as, Cy3.TM., as the reporter linked to the 5' end of a reporting DNA sequence. The second method positions a reporter dye, e.g., FAM, on the 3' end of the reporting DNA sequence and a quencher dye, e.g., TAMRA, on the 5' end.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue Aug 01 00:00:00 EDT 2006},
month = {Tue Aug 01 00:00:00 EDT 2006}
}

Works referenced in this record:

A Reusable Flow-Through Polymerase Chain Reaction Instrument for the Continuous Monitoring of Infectious Biological Agents
journal, July 2003


Multiplexed single nucleotide polymorphism genotyping by oligonucleotide ligation and flow cytometry
journal, February 2000


Classification and properties of 64 multiplexed microsphere sets
journal, October 1998


A handheld real time thermal cycler for bacterial pathogen detection
journal, August 2003


Advanced multiplexed analysis with the FlowMetrixTM system
journal, September 1997


Flow Cytometric Platform for High-Throughput Single Nucleotide Polymorphism Analysis
journal, March 2001


Suspension arrays for high throughput, multiplexed single nucleotide polymorphism genotyping
journal, June 2000


A multiplexed PCR-coupled liquid bead array for the simultaneous detection of four biothreat agents
journal, April 2005


Autonomous Detection of Aerosolized Biological Agents by Multiplexed Immunoassay with Polymerase Chain Reaction Confirmation
journal, January 2005