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

Patent · Tue Sep 03 00:00:00 EDT 1996

OSTI ID:912997

Khrapko, Konstantin R ^[1]; Khorlin, Alexandr A ^[1]; Ivanov, Igor B ^[2]; Ershov, Gennady M ^[1]; Lysov, Jury P ^[1]; Florentiev, Vladimir L ^[1]; Mirzabekov, Andrei D ^[1]

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1. Moscow, RU
2. Moskovskaya, RU

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 (United States)
• Sponsoring Organization: USDOE
• 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

Related Subjects

59 BASIC BIOLOGICAL SCIENCES