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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.TM. on the 3' end of the reporting DNA sequence and a quencher dye, e.g., TAMRA.TM., on the 5' end.

Inventors:
 [1];  [1];  [2]
  1. Livermore, CA
  2. Round Rock, TX
Issue Date:
Research Org.:
Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1019570
Patent Number(s):
7972818
Application Number:
US Patent Application 11/454,478
Assignee:
Lawrence Livermore National Security, LLC (Livermore, 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., 2011. 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/1019570.
@article{osti_1019570,
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.TM. on the 3' end of the reporting DNA sequence and a quencher dye, e.g., TAMRA.TM., on the 5' end.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2011},
month = {7}
}

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Works referenced in this record:

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


Detection of specific polymerase chain reaction product by utilizing the 5'----3' exonuclease activity of Thermus aquaticus DNA polymerase.
journal, August 1991


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


A Bead-Based Method for Multiplexed Identification and Quantitation of DNA Sequences Using Flow Cytometry
journal, October 2000


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