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Title: Methods for analyzing nucleic acid sequences

Abstract

The present invention is directed to a method of sequencing a target nucleic acid. The method provides a complex comprising a polymerase enzyme, a target nucleic acid molecule, and a primer, wherein the complex is immobilized on a support Fluorescent label is attached to a terminal phosphate group of the nucleotide or nucleotide analog. The growing nucleic acid strand is extended by using the polymerase to add a nucleotide analog to the nucleic acid strand. The nucleotide analog added to the oligonucleotide primer as a result of the polymerizing step is identified. The time duration of the signal from labeled nucleotides or nucleotide analogs that become incorporated is distinguished from freely diffusing labels by a longer retention in the observation volume for the nucleotides or nucleotide analogs that become incorporated than for the freely diffusing labels.

Inventors:
 [1];  [1];  [1];  [1];  [1];  [1]
  1. Ithaca, NY
Issue Date:
Research Org.:
Cornell Research Foundation (Ithaca, NY)
Sponsoring Org.:
USDOE
OSTI Identifier:
1020255
Patent Number(s):
7943307
Application Number:
11/336,122
Assignee:
Cornell Research Foundation (Ithaca, NY)
Patent Classifications (CPCs):
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
Y - NEW / CROSS SECTIONAL TECHNOLOGIES Y10 - TECHNICAL SUBJECTS COVERED BY FORMER USPC Y10S - TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
DOE Contract Number:  
066898-0003891
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Korlach, Jonas, Webb, Watt W, Levene, Michael, Turner, Stephen, Craighead, Harold G, and Foquet, Mathieu. Methods for analyzing nucleic acid sequences. United States: N. p., 2011. Web.
Korlach, Jonas, Webb, Watt W, Levene, Michael, Turner, Stephen, Craighead, Harold G, & Foquet, Mathieu. Methods for analyzing nucleic acid sequences. United States.
Korlach, Jonas, Webb, Watt W, Levene, Michael, Turner, Stephen, Craighead, Harold G, and Foquet, Mathieu. Tue . "Methods for analyzing nucleic acid sequences". United States. https://www.osti.gov/servlets/purl/1020255.
@article{osti_1020255,
title = {Methods for analyzing nucleic acid sequences},
author = {Korlach, Jonas and Webb, Watt W and Levene, Michael and Turner, Stephen and Craighead, Harold G and Foquet, Mathieu},
abstractNote = {The present invention is directed to a method of sequencing a target nucleic acid. The method provides a complex comprising a polymerase enzyme, a target nucleic acid molecule, and a primer, wherein the complex is immobilized on a support Fluorescent label is attached to a terminal phosphate group of the nucleotide or nucleotide analog. The growing nucleic acid strand is extended by using the polymerase to add a nucleotide analog to the nucleic acid strand. The nucleotide analog added to the oligonucleotide primer as a result of the polymerizing step is identified. The time duration of the signal from labeled nucleotides or nucleotide analogs that become incorporated is distinguished from freely diffusing labels by a longer retention in the observation volume for the nucleotides or nucleotide analogs that become incorporated than for the freely diffusing labels.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2011},
month = {5}
}

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Works referenced in this record:

Near-Field Fluorescence Microscopy Based on Two-Photon Excitation with Metal Tips
journal, May 1999


Rapid and simple preparation of plasmids suitable for dideoxy DNA sequencing and other purposes
journal, January 1991


Dual-color fluorescence cross-correlation spectroscopy for multicomponent diffusional analysis in solution
journal, April 1997


Probing Single Molecule Dynamics
journal, July 1994


Solid Phase DNA Minisequencing by an Enzymatic Luminometric Inorganic Pyrophosphate Detection Assay
journal, January 1993


Rapid DNA sequencing based upon single molecule detection
journal, February 1991


Single-Molecule Fluorescence Analysis in Solution
journal, July 1996


DNA sequencing with chain-terminating inhibitors
journal, December 1977


Single molecule fluorescence: From excitation spectra to time correlation
journal, April 1994


Single-Molecule Detection of Specific Nucleic Acid Sequences in Unamplified Genomic DNA
journal, October 1997


Real-Time DNA Sequencing Using Detection of Pyrophosphate Release
journal, November 1996


Fluorescence Spectroscopy of Single Biomolecules
journal, March 1999


Fluorescence of Intramolecular and Intermolecular Interactions of Aminonaphthyl-sulfonate with Nucleotides
journal, August 1982


Sensitized Photomodification of Single-Stranded DNA by a Binary System of Oligonucleotide Conjugates
journal, August 1997


Single-molecule fluorescence spectroscopy of enzyme conformational dynamics and cleavage mechanism
journal, February 1999


Alterations of Single Molecule Fluorescence Lifetimes in Near-Field Optical Microscopy
journal, July 1994


Theory of Nanometric Optical Tweezers
journal, July 1997


Single-molecule detection as an approach to rapid DNA sequencing
journal, January 1992


Monitoring conformational dynamics of a single molecule by selective fluorescence spectroscopy
journal, February 1998


Three-Dimensional Imaging of Single Molecules Solvated in Pores of Poly(acrylamide) Gels
journal, November 1996


Feasibility of molecular-resolution fluorescence near-field microscopy using multi-photon absorption and field enhancement near a sharp tip
journal, February 1999


Application of Single Molecule Detection to Dna Sequencing
journal, May 1997