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Title: Method for the detection of specific nucleic acid sequences by polymerase nucleotide incorporation

Abstract

A method for rapid and efficient detection of a target DNA or RNA sequence is provided. A primer having a 3'-hydroxyl group at one end and having a sequence of nucleotides sufficiently homologous with an identifying sequence of nucleotides in the target DNA is selected. The primer is hybridized to the identifying sequence of nucleotides on the DNA or RNA sequence and a reporter molecule is synthesized on the target sequence by progressively binding complementary nucleotides to the primer, where the complementary nucleotides include nucleotides labeled with a fluorophore. Fluorescence emitted by fluorophores on single reporter molecules is detected to identify the target DNA or RNA sequence.

Inventors:
Publication Date:
Research Org.:
The Regents of the University of California, Los Alamos, NM (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1174871
Patent Number(s):
6,743,578
Application Number:
09/454,385
Assignee:
University Of California, The Regents Of OSTI
DOE Contract Number:
W-7405-ENG-36
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Castro, Alonso. Method for the detection of specific nucleic acid sequences by polymerase nucleotide incorporation. United States: N. p., 2004. Web.
Castro, Alonso. Method for the detection of specific nucleic acid sequences by polymerase nucleotide incorporation. United States.
Castro, Alonso. Tue . "Method for the detection of specific nucleic acid sequences by polymerase nucleotide incorporation". United States. doi:. https://www.osti.gov/servlets/purl/1174871.
@article{osti_1174871,
title = {Method for the detection of specific nucleic acid sequences by polymerase nucleotide incorporation},
author = {Castro, Alonso},
abstractNote = {A method for rapid and efficient detection of a target DNA or RNA sequence is provided. A primer having a 3'-hydroxyl group at one end and having a sequence of nucleotides sufficiently homologous with an identifying sequence of nucleotides in the target DNA is selected. The primer is hybridized to the identifying sequence of nucleotides on the DNA or RNA sequence and a reporter molecule is synthesized on the target sequence by progressively binding complementary nucleotides to the primer, where the complementary nucleotides include nucleotides labeled with a fluorophore. Fluorescence emitted by fluorophores on single reporter molecules is detected to identify the target DNA or RNA sequence.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue Jun 01 00:00:00 EDT 2004},
month = {Tue Jun 01 00:00:00 EDT 2004}
}

Patent:

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  • A method is detected for detecting the presence or absence of at least one specific restriction site in a specific nucleic acid sequence comprising the steps of: (a) hybridizing the nucleic acid sequence in solution with an oligonucleotide probe for each restriction site detected. A probe is complementary to a region in the nucleic acid sequence spanning the respective restriction site of the probe is labeled at the end which is nearer to the respective restriction site than the other end of the probe; (b) digesting the hybridized nucleic acid sequence with a restriction endonuclease for each restriction site detectedmore » by each probe capable of cleaving its respective probe at the restriction site being detected. This produces labeled and unlabeled oligomer fragments.; (c) separating any labeled cleaved oligomer fragments from labeled uncleaved oligomers, and (d) detecting the presence or absence of labeled oligomer fragments.« less
  • Nucleotide sequences specific to Yersinia pestis that serve as markers or signatures for identification of this bacterium were identified. In addition, forward and reverse primers and hybridization probes derived from these nucleotide sequences that are used in nucleotide detection methods to detect the presence of the bacterium are disclosed.
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  • Nucleotide sequences specific to Brucella that serves as a marker or signature for identification of this bacterium were identified. In addition, forward and reverse primers and hybridization probes derived from these nucleotide sequences that are used in nucleotide detection methods to detect the presence of the bacterium are disclosed.
  • Described herein is the identification of nucleotide sequences specific to Francisella tularensis that serves as a marker or signature for identification of this bacterium. In addition, forward and reverse primers and hybridization probes derived from these nucleotide sequences that are used in nucleotide detection methods to detect the presence of the bacterium are disclosed.