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Title: Method for sequencing nucleic acid molecules

Abstract

The present invention is directed to a method of sequencing a target nucleic acid molecule having a plurality of bases. In its principle, the temporal order of base additions during the polymerization reaction is measured on a molecule of nucleic acid, i.e. the activity of a nucleic acid polymerizing enzyme on the template nucleic acid molecule to be sequenced is followed in real time. The sequence is deduced by identifying which base is being incorporated into the growing complementary strand of the target nucleic acid by the catalytic activity of the nucleic acid polymerizing enzyme at each step in the sequence of base additions. A polymerase on the target nucleic acid molecule complex is provided in a position suitable to move along the target nucleic acid molecule and extend the oligonucleotide primer at an active site. A plurality of labelled types of nucleotide analogs are provided proximate to the active site, with each distinguishable type of nucleotide analog being complementary to a different nucleotide in the target nucleic acid sequence. The growing nucleic acid strand is extended by using the polymerase to add a nucleotide analog to the nucleic acid strand at the active site, where the nucleotide analog beingmore » added is complementary to the nucleotide of the target nucleic acid at the active site. The nucleotide analog added to the oligonucleotide primer as a result of the polymerizing step is identified. The steps of providing labelled nucleotide analogs, polymerizing the growing nucleic acid strand, and identifying the added nucleotide analog are repeated so that the nucleic acid strand is further extended and the sequence of the target nucleic acid is determined.« less

Inventors:
; ; ; ; ;
Issue Date:
Research Org.:
Cornell Research Foundation, Inc., Ithaca, NY (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1175760
Patent Number(s):
7,052,847
Application Number:
11/013,578
Assignee:
Cornell Research Foundation, Inc. (Ithaca, NY)
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. Method for sequencing nucleic acid molecules. United States: N. p., 2006. Web.
Korlach, Jonas, Webb, Watt W., Levene, Michael, Turner, Stephen, Craighead, Harold G., & Foquet, Mathieu. Method for sequencing nucleic acid molecules. United States.
Korlach, Jonas, Webb, Watt W., Levene, Michael, Turner, Stephen, Craighead, Harold G., and Foquet, Mathieu. Tue . "Method for sequencing nucleic acid molecules". United States. https://www.osti.gov/servlets/purl/1175760.
@article{osti_1175760,
title = {Method for sequencing nucleic acid molecules},
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 molecule having a plurality of bases. In its principle, the temporal order of base additions during the polymerization reaction is measured on a molecule of nucleic acid, i.e. the activity of a nucleic acid polymerizing enzyme on the template nucleic acid molecule to be sequenced is followed in real time. The sequence is deduced by identifying which base is being incorporated into the growing complementary strand of the target nucleic acid by the catalytic activity of the nucleic acid polymerizing enzyme at each step in the sequence of base additions. A polymerase on the target nucleic acid molecule complex is provided in a position suitable to move along the target nucleic acid molecule and extend the oligonucleotide primer at an active site. A plurality of labelled types of nucleotide analogs are provided proximate to the active site, with each distinguishable type of nucleotide analog being complementary to a different nucleotide in the target nucleic acid sequence. The growing nucleic acid strand is extended by using the polymerase to add a nucleotide analog to the nucleic acid strand at the active site, where the nucleotide analog being added is complementary to the nucleotide of the target nucleic acid at the active site. The nucleotide analog added to the oligonucleotide primer as a result of the polymerizing step is identified. The steps of providing labelled nucleotide analogs, polymerizing the growing nucleic acid strand, and identifying the added nucleotide analog are repeated so that the nucleic acid strand is further extended and the sequence of the target nucleic acid is determined.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2006},
month = {5}
}

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