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Method of sequencing by hybridization of oligonucleotide probes

Patent 5525464 Issued on June 11, 1996. Estimated Expiration Date: Icon_subject February 28, 2014. Estimated Expiration Date is calculated based on simple USPTO term provisions. It does not account for terminal disclaimers, term adjustments, failure to pay maintenance fees, or other factors which might affect the term of a patent.

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Inventors

Application

No. 203502 filed on 02/28/1994

US Classes:

435/6, Involving nucleic acid435/91.1, Polynucleotide (e.g., nucleic acid, oligonucleotide, etc.)435/91.2Acellular exponential or geometric amplification (e.g., PCR, etc.)

Examiners

Primary: Fleisher, Mindy
Assistant: Ketter, James

Attorney, Agent or Firm

Foreign Patent References

  • 0152886 EP. 08/13/1985
  • 3506703 DE. 10/13/1986
  • 88/01302 WO. 02/13/1988
  • 89/10977 WO. 11/13/1989
  • 90/03382 WO. 04/13/1990
  • 90/04652 WO. 05/13/1990

International Class

C12Q 001/68

Foreign Application Priority Data

1987-04-01 YU

Abstract

The conditions under which oligonucleotide probes hybridize preferentially with entirely complementary and homologous nucleic acid targets are described. Using these hybridization conditions, overlapping oligonucleotide probes associate with a target nucleic acid. Following washes, positive hybridization signals are used to assemble the sequence of a given nucleic acid fragment. Representative target nucleic acids are applied as dots. Up to to 100,000 probes of the type (A,T,C,G)(A,T,C,G)N8(A,T,C,G) are used to determine sequence information by simultaneous hybridization with nucleic acid molecules bound to a filter. Additional hybridization conditions are provided that allow stringent hybridization of 6-10 nucleotide long oligomers which extends the utility of the invention. A computer process determines the information sequence of the target nucleic acid which can include targets with the complexity of mammalian genomes. Sequence generation can be obtained for a large complex mammalian genome in a single process.

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