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US5104792A Expired / Lapsed US ⏱️ 0.0 years remaining

Method for amplifying unknown nucleic acid sequences

Assignee: United States Department of Health and Human Services
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Filing: 1989-12-21
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Granted: 1992-04-14
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Expires: 2009-12-21
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Abstract

Abstract A modification of the PCR technique is described which allows fragments of RNA or DNA to be amplified without prior knowledge of their sequence. The technique can be used to amplify viral nucleic acids present in small amounts in clinical material allowing, for example, the diagnosis of a particular virus infection or the discovery of new viruses.

Inventors

  • 👤 Jonathan Silver
  • 👤 Stephen Feinstone

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-10-29 22:22
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6844; C12Q1/6853; C12Q1/686

Claims Analysis

Total Claims: 2
Claim 1
Independent Claim 139 words
A method for performing polymerase chain reaction (PCR) on nucleic acid fragments of unknown sequence, comprising the sequential steps of: (a) obtaining purified template RNA or DNA to be amplified; (b) annealing to template RNA or DNA universal primers whose 5' ends have a fixed sequence of sufficient length to serve as a primer in a subsequent polymerase chain reaction, and whose 3' ends are degenerate for random priming of the template; (c) extending the universal primers by two or more rounds of primer extension with a polymerase capable of copying a RNA or a DNA template; (d) removing the 3' degenerate oligonucleotides; (e) amplifying with PCR using primers whose 3' sequence is the same as the fixed 5' portion of the universal oligonucleotides; (f) then identifying the PCR product obtained from step (e) by standard molecular biological techniques.
Claim 2
Independent Claim 139 words
A method for performing polymerase chain reaction (PCR) on nucleic acid fragments of unknown sequence, comprising the sequential steps of: (a) obtaining purified template RNA or DNA to be amplified; (b) annealing to template RNA or DNA universal primers whose 5' ends have a fixed sequence of sufficient length to serve as a primer in a subsequent polymerase chain reaction, and whose 3' ends are degenerate for random priming of the template; (c) extending the universal primers by two or more rounds of primer extension with a polymerase capable of copying a RNA or a DNA template; (d) removing the 3' degenerate oligonucleotides; (e) amplifying with PCR using primers whose 3' sequence is the same as the fixed 5' portion of the universal oligonucleotides; (f) then identifying the PCR product obtained from step (e) by standard molecular biological techniques.

References Cited (Backward Citations)

EP0224126A2 Cited
US4683195A Cited
US4683195B1 Cited

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1995-04-14 1995-10-14 1996-04-14 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1999-04-14 1999-10-14 2000-04-14 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 2003-04-14 2003-10-14 2004-04-14 $7700.00 Window Closed / Lapsed

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