US5521065A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Detection of specific sequences in nucleic acids
Assignee: Applied Biosystems LLC
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Filing: 1994-06-08
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Granted: 1996-05-28
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Expires: 2014-06-08
Abstract
Abstract A method of testing for the presence or absence of a target sequence in a mixture of single-stranded nucleic acid fragments is disclosed. The method involves reacting a mixture of single-stranded nucleic acid fragments with a first probe which is complementary to a first region of the target sequence, and with a second probe which is complementary to a second region of the target sequence, where the first and second target regions are contiguous with one another, under hybridization conditions in which the two probes become stably hybridized to their associated target regions. Following hybridization, any of the first and second probes hybridized to contiguous first and second target regions are ligated, and the sample is tested for the presence of expected probe ligation product. The presence of ligated product indicates that the target sequence is present in the sample.
Inventors
- 👤 Norman M. Whiteley
- 👤 Michael W. Hunkapiller
- 👤 Alexander N. Glazer
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 12:09
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6844; C12Q1/6862; C12Q1/6813; C12Q1/6827; Y; Y10; Y10S; Y10S435/00; Y10S435/803; Y10S435/81
Claims Analysis
Total Claims: 8
Claim 1
Independent Claim
139 words
A method of testing for the presence or absence of a target sequence in a mixture of single-stranded nucleic acid fragments, comprising: reacting a mixture of single-stranded nucleic acid fragments with a first probe which is complementary to a first region of the target sequence, and with a second probe which is complementary to a second region of the target sequence, where the first and second target regions are contiguous with one another, under hybridization conditions in which the two probes become stably hybridized to their associated target regions, ligating first and second probes which have hybridized to said contiguous first and second target regions, and testing for the presence of ligated first and second probes, where the presence of the ligation product of said first and second probes indicates the presence of the target sequence in said mixture.
Claim 2
Independent Claim
139 words
A method of testing for the presence or absence of a target sequence in a mixture of single-stranded nucleic acid fragments, comprising: reacting a mixture of single-stranded nucleic acid fragments with a first probe which is complementary to a first region of the target sequence, and with a second probe which is complementary to a second region of the target sequence, where the first and second target regions are contiguous with one another, under hybridization conditions in which the two probes become stably hybridized to their associated target regions, ligating first and second probes which have hybridized to said contiguous first and second target regions, and testing for the presence of ligated first and second probes, where the presence of the ligation product of said first and second probes indicates the presence of the target sequence in said mixture.
Claim 3
Dependent Claim
15 words
The method of claim 1, wherein one of the first and second probes is labeled.
Claim 4
Dependent Claim
26 words
The method of claim 2, wherein said testing includes separating labeled, ligated probes from labeled, non-ligated probes, and assaying for the presence of labeled, ligated probes.
Claim 5
Dependent Claim
21 words
The method of claim 1, wherein said testing further includes dissociating ligated probe from the target sequence prior to said testing.
Claim 6
Dependent Claim
15 words
The method of claim 1, wherein said ligating is carried out using a ligase enzyme.
Claim 7
Dependent Claim
15 words
The method of claim 1, wherein the target sequence tested for contains a genetic mutation.
Claim 8
Dependent Claim
33 words
The method of claim 1, wherein said nucleic acid mixture is tested for the presence of a normal target sequence and for the presence of a corresponding target sequence containing a genetic mutation.
References Cited (Backward Citations)
US4563419A
Cited
US4883750A
Cited
US5242794A
Cited
US5093232A
Cited
US4925785A
Cited
US5082830A
Cited
WO1989009835A1
Cited
US5109124A
Cited
US5185243A
Cited
EP0439182B1
Cited
US5138045A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1999-05-28 | 1999-11-27 | 2000-05-28 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2003-05-28 | 2003-11-27 | 2004-05-28 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2007-05-28 | 2007-11-27 | 2008-05-28 | $7700.00 | Window Closed / Lapsed |