US5763181A
Expired / Lapsed
US
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Continous fluorometric assay for detecting nucleic acid cleavage
Assignee: Georgetown University
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Filing: 1996-08-30
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Granted: 1998-06-09
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Expires: 2016-08-30
Abstract
Abstract A method of detecting an enzyme-mediated DNA cleavage reaction in a fluorometric assay is provided. The method can be used to detect DNA cleavage caused by restriction endonucleases, retroviral integrase enzymes, DNases, RNases, or enzymes utilized in other strand separating processes in molecular biology.
Inventors
- 👤 Myun Ki Han
- 👤 S. Paul Lee
- 👤 Jack G. Chirikjian
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 07:10
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6816; C12Q1/6818; C12Q1/34; C12Q1/44; C12Q1/6823
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
83 words
A method of detecting continuously an endonuclease-mediated nucleic acid cleavage reaction in a fluorometric assay comprising the steps of: (a) preparing a fluorescently labeled oligonucleotide containing a double-stranded nucleic acid sequence recognizable by said endonuclease, wherein said oligonucleotide acts as a substrate for said endonuclease; (b) contacting said oligonucleotide of step (a) with said endonuclease in an amount sufficient to cleave said oligonucleotide to produce oligonucleotide products; and (c) detecting continuously a nucleic acid cleavage reaction by detecting an increase in fluorescence intensity.
Claim 2
Independent Claim
83 words
A method of detecting continuously an endonuclease-mediated nucleic acid cleavage reaction in a fluorometric assay comprising the steps of: (a) preparing a fluorescently labeled oligonucleotide containing a double-stranded nucleic acid sequence recognizable by said endonuclease, wherein said oligonucleotide acts as a substrate for said endonuclease; (b) contacting said oligonucleotide of step (a) with said endonuclease in an amount sufficient to cleave said oligonucleotide to produce oligonucleotide products; and (c) detecting continuously a nucleic acid cleavage reaction by detecting an increase in fluorescence intensity.
Claim 3
Dependent Claim
14 words
The method of claim 1, wherein said oligonucleotide is fluorescently labeled at one end.
Claim 4
Dependent Claim
14 words
The method of claim 1, wherein said oligonucleotide is fluorescently labeled at both ends.
Claim 5
Dependent Claim
11 words
The method of claim 1, wherein said nucleic acid is DNA.
Claim 6
Dependent Claim
12 words
The method of claim 1, wherein said endonuclease is a restriction endonuclease.
Claim 7
Dependent Claim
14 words
The method of claim 1, wherein said endonuclease is a DNase or RNase enzyme.
Claim 8
Dependent Claim
13 words
The method of claim 1, wherein said endonuclease is a retroviral integrase enzyme.
Claim 9
Dependent Claim
12 words
The method of claim 2, wherein said endonuclease is a restriction endonuclease.
Claim 10
Dependent Claim
13 words
The method of claim 3, wherein said endonuclease is a retroviral integrase enzyme.
Claim 11
Dependent Claim
11 words
The method of claim 5, wherein said restriction endonuclease is BamHI.
Claim 12
Dependent Claim
13 words
The method of claim 7, wherein said retroviral integrase enzyme is HIV integrase.
Claim 13
Dependent Claim
26 words
The method of claim 1, wherein said endonuclease-mediated nucleic acid cleavage reaction occurs during a process for amplifying or detecting a specific DNA or RNA sequence.
Claim 14
Dependent Claim
21 words
The method of claim 12, wherein said process for amplifying or detecting a DNA or RNA sequence is catalytic hybridization amplification.
Claim 15
Dependent Claim
11 words
The method of claim 13, wherein said endonuclease is RNase H.
Claim 16
Dependent Claim
14 words
The method of claim 14, wherein said oligonucleotide is fluorescently labeled at both ends.
References Cited (Backward Citations)
US4683194A
Cited
US4683202A
Cited
US4683202B1
Cited
US4876187A
Cited
US4996143A
Cited
US4683195A
Cited
US4683195B1
Cited
WO1989009284A1
Cited
WO1989009835A1
Cited
US5001769A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2001-06-08 | 2001-12-08 | 2002-06-09 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2005-06-08 | 2005-12-08 | 2006-06-09 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2009-06-08 | 2009-12-08 | 2010-06-09 | $7700.00 | Window Closed / Lapsed |