skip to main content
DOE Patents title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Methods of DNA sequencing by hybridization based on optimizing concentration of matrix-bound oligonucleotide and device for carrying out same

Abstract

A method for sequencing DNA by hybridization that includes the following steps: forming an array of oligonucleotides at such concentrations that either ensure the same dissociation temperature for all fully complementary duplexes or allows hybridization and washing of such duplexes to be conducted at the same temperature; hybridizing said oligonucleotide array with labeled test DNA; washing in duplex dissociation conditions; identifying single-base substitutions in the test DNA by analyzing the distribution of the dissociation temperatures and reconstructing the DNA nucleotide sequence based on the above analysis. A device for carrying out the method comprises a solid substrate and a matrix rigidly bound to the substrate. The matrix contains the oligonucleotide array and consists of a multiplicity of gel portions. Each gel portion contains one oligonucleotide of desired length. The gel portions are separated from one another by interstices and have a thickness not exceeding 30 .mu.m.

Inventors:
 [1];  [1];  [2];  [1];  [1];  [1];  [1]
  1. Moscow, RU
  2. Moskovskaya, RU
Issue Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
912997
Patent Number(s):
5552270
Assignee:
Institut Molekulyarnoi Biologii Imeni V.A. (Moscow, RU)
Patent Classifications (CPCs):
B - PERFORMING OPERATIONS B01 - PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL B01J - CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Khrapko, Konstantin R, Khorlin, Alexandr A, Ivanov, Igor B, Ershov, Gennady M, Lysov, Jury P, Florentiev, Vladimir L, and Mirzabekov, Andrei D. Methods of DNA sequencing by hybridization based on optimizing concentration of matrix-bound oligonucleotide and device for carrying out same. United States: N. p., 1996. Web.
Khrapko, Konstantin R, Khorlin, Alexandr A, Ivanov, Igor B, Ershov, Gennady M, Lysov, Jury P, Florentiev, Vladimir L, & Mirzabekov, Andrei D. Methods of DNA sequencing by hybridization based on optimizing concentration of matrix-bound oligonucleotide and device for carrying out same. United States.
Khrapko, Konstantin R, Khorlin, Alexandr A, Ivanov, Igor B, Ershov, Gennady M, Lysov, Jury P, Florentiev, Vladimir L, and Mirzabekov, Andrei D. Tue . "Methods of DNA sequencing by hybridization based on optimizing concentration of matrix-bound oligonucleotide and device for carrying out same". United States. https://www.osti.gov/servlets/purl/912997.
@article{osti_912997,
title = {Methods of DNA sequencing by hybridization based on optimizing concentration of matrix-bound oligonucleotide and device for carrying out same},
author = {Khrapko, Konstantin R and Khorlin, Alexandr A and Ivanov, Igor B and Ershov, Gennady M and Lysov, Jury P and Florentiev, Vladimir L and Mirzabekov, Andrei D},
abstractNote = {A method for sequencing DNA by hybridization that includes the following steps: forming an array of oligonucleotides at such concentrations that either ensure the same dissociation temperature for all fully complementary duplexes or allows hybridization and washing of such duplexes to be conducted at the same temperature; hybridizing said oligonucleotide array with labeled test DNA; washing in duplex dissociation conditions; identifying single-base substitutions in the test DNA by analyzing the distribution of the dissociation temperatures and reconstructing the DNA nucleotide sequence based on the above analysis. A device for carrying out the method comprises a solid substrate and a matrix rigidly bound to the substrate. The matrix contains the oligonucleotide array and consists of a multiplicity of gel portions. Each gel portion contains one oligonucleotide of desired length. The gel portions are separated from one another by interstices and have a thickness not exceeding 30 .mu.m.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1996},
month = {9}
}

Patent:

Save / Share:

Works referenced in this record:

Improved Chips for Sequencing by Hybridization
journal, October 1991


Detection of single base changes in nucleic acids
journal, October 1989


A method for DNA sequencing by hybridization with oligonucleotide matrix
journal, January 1991


An oligonucleotide hybridization approach to DNA sequencing
journal, October 1989


DNA sequencing by hybridization: 100 bases read by a non-gel-based method.
journal, November 1991


Dissociation of Duplexes Formed by Hybridization of DNA with Gel-Immobilized Oligonucleotides
journal, February 1994


A novel method for nucleic acid sequence determination
journal, December 1988


1-Tuple DNA Sequencing: Computer Analysis
journal, August 1989