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:

Korlach, Jonas ^[1]; Webb, Watt W ^[1]; Levene, Michael ^[1]; Turner, Stephen ^[1]; Craighead, Harold G ^[1]; Foquet, Mathieu ^[1]

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+ Show Inventor Affiliations

1. Ithaca, NY

Issue Date:

Tue May 17 00:00:00 EDT 2011

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

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C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS

C12Q1/6869 - Methods for sequencing
C12Q1/6874 - involving nucleic acid arrays, e.g. sequencing by hybridisation
C12Q2521/543 - Immobilised enzyme
C12Q2537/149 - Sequential reactions
C12Q2561/113 - Real time assay
C12Q2561/12 - Fluorescence lifetime measurement
C12Q2565/518 - characterised by the immobilisation of the nucleic acid sample or target
C12Q2565/537 - characterised by the capture oligonucleotide acting as a primer

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

Y10S436/80 - Fluorescent dyes, e.g. rhodamine
Y10S436/805 - Optical property

Y - NEW / CROSS SECTIONAL TECHNOLOGIES Y10 - TECHNICAL SUBJECTS COVERED BY FORMER USPC Y10T - TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

Y10T436/143333 - Saccharide [e.g., DNA, etc.]

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DOE Contract Number:  

066898-0003891

Resource Type:

Patent

Country of Publication:

United States

Language:

English

Subject:

59 BASIC BIOLOGICAL SCIENCES