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

Patent ·
OSTI ID:912997
 [1];  [1];  [2];  [1];  [1];  [1];  [1]
  1. Moscow, RU
  2. Moskovskaya, RU
+ Show Author Affiliations
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.
Research Organization:
Argonne National Laboratory (ANL), Argonne, IL
Sponsoring Organization:
United States Department of Energy
Assignee:
Institut Molekulyarnoi Biologii Imeni V.A. (Moscow, RU)
Patent Number(s):
5,552,270
OSTI ID:
912997
Country of Publication:
United States
Language:
English

References (9)

A novel method for nucleic acid sequence determination journal December 1988
1-Tuple DNA Sequencing: Computer Analysis journal August 1989
Analyzing and comparing nucleic acid sequences by hybridization to arrays of oligonucleotides: Evaluation using experimental models journal August 1992
Dissociation of Duplexes Formed by Hybridization of DNA with Gel-Immobilized Oligonucleotides journal February 1994
A method for DNA sequencing by hybridization with oligonucleotide matrix journal January 1991
Detection of single base changes in nucleic acids journal October 1989
DNA sequencing by hybridization: 100 bases read by a non-gel-based method. journal November 1991
Improved Chips for Sequencing by Hybridization journal October 1991
An oligonucleotide hybridization approach to DNA sequencing journal October 1989

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES