US5030557A
Expired / Lapsed
US
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Means and method for enhancing nucleic acid hybridization
Assignee: Gen Probe Inc
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Filing: 1987-11-24
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Granted: 1991-07-09
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Expires: 2007-11-24
Abstract
Abstract The binding of a nucleic acid probe with its complementary sequence in a targeted, single stranded nucleic acid is affected by the secondary and tertiary structure of the target nucleic acid. The rate and extent of hybridization of the probe with the targeted nucleic acid can be increased by the use of "helper" oligonucleotides. Helper oligonucleotides are selected to bind to the target nucleic acid and impose a different secondary and tertiary structure on the target to facilitate the binding of the probe to the target. The resulting hybrid of probe and target nucleic acid also exhibits a higher T m than the hybrid which results from addition of the probe alone.
Inventors
- 👤 James J. Hogan
- 👤 Curt L. Milliman
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 16:47
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/02; C12Q1/04; C12Q1/12; C12Q1/6813; C12Q1/6832; C12Q1/6876; C12Q1/6888; C12Q1/689; Y; Y10; Y10S; Y10S435/00; Y10S435/81
Claims Analysis
Total Claims: 30
Claim 1
Independent Claim
67 words
A process for enhancing the binding between a nucleotide probe and a complementary nucleotide sequence in a single stranded target nucleic acid comprising adding to the target nucleic acid a helper oligonucleotide which hybridizes with the target nucleic acid in a different region than the probe, the helper oligonucleotide being added in an amount effective to enhance the binding of the probe to the target nucleic acid.
Claim 2
Dependent Claim
67 words
A process for enhancing the binding between a nucleotide probe and a complementary nucleotide sequence in a single stranded target nucleic acid comprising adding to the target nucleic acid a helper oligonucleotide which hybridizes with the target nucleic acid in a different region than the probe, the helper oligonucleotide being added in an amount effective to enhance the binding of the probe to the target nucleic acid.
Claim 3
Dependent Claim
19 words
A process according to claim 1 wherein the target nucleic acid is selected from DNA, mRNA, rRNA and tRNA.
Claim 4
Dependent Claim
36 words
A process according to claim 2 wherein the target nucleic acid is separated from an organism producing it or in which it is found prior to the addition of the helper oligonucleotide and the nucleotide probe.
Claim 5
Dependent Claim
44 words
A process according to claim 2 wherein the helper oligonucleotide and the nucleotide probe are introduced to a cell in which the target nucleic acid is located and the hybridization between the target nucleic acid and the helper oligonucleotide and nucleotide probe occurs intracellularly.
Claim 6
Dependent Claim
27 words
A process according to any of claims 1, 2, 3 or 4 wherein the probe is a DNA oligonucleotide and the helper oligonucleotide is a DNA oligonucleotide.
Claim 7
Dependent Claim
33 words
A process according to claim 5 wherein the probe comprises from about 10 to about 50 nucleotides in length and the helper oligonucleotides comprise from about 10 to about 100 nucleotides in length.
Claim 8
Dependent Claim
30 words
A process according to claim 6 wherein the probe and helper oligonucleotides are selected from DNA having a diphosphate ester backbone, an alkyl or arylphosphonate backbone or a phosphorothioate backbone.
Claim 9
Dependent Claim
28 words
A process according to claim 7 wherein the probe comprises about 15 to about 40 nucleotides and the helper oligonucleotides comprise from about 20 to about 50 nucleotides.
Claim 10
Dependent Claim
21 words
A process according to claim 6 wherein the helper oligonucleotide binds with the target nucleic acid immediately adjacent to the probe.
Claim 11
Dependent Claim
21 words
A process according to claim 7 wherein the helper oligonucleotide binds with the target nucleic acid immediately adjacent to the probe.
Claim 12
Dependent Claim
23 words
A process according to claim 6 wherein the helper oligonucleotide binds with the target nucleic acid in a region removed from the probe.
Claim 13
Dependent Claim
23 words
A process according to claim 7 wherein the helper oligonucleotide binds with the target nucleic acid in a region remote from the probe.
Claim 14
Dependent Claim
18 words
A process according to claim 5 wherein the probe is labeled to permit the detection of the probe.
Claim 15
Dependent Claim
21 words
A process according to claim 13 wherein the probe is detected after formation of hybrid and separation from the unhybridized probe.
Claim 16
Dependent Claim
28 words
A process according to claim 14 wherein the label is a radionucleotide, an enzyme, a fluorometric moiety or a moiety which participates in a reaction which produces chemiluminescence.
Claim 17
Dependent Claim
18 words
A process according to claim 15 wherein the label is elected from 125 I or an acridinium ester.
Claim 18
Dependent Claim
14 words
A process according to claim 5 wherein a mixture of helper oligonucleotides is used.
Claim 19
Dependent Claim
14 words
A process according to claim 6 wherein a mixture of helper oligonucleotides is used.
Claim 20
Dependent Claim
14 words
A process according to claim 14 wherein a mixture of helper oligonucleotides is used.
Claim 21
Dependent Claim
14 words
A process according to claim 15 wherein a mixture of helper oligonucleotides is used.
Claim 22
Dependent Claim
23 words
A hybrid nucleic acid comprising a hybrid of a nucleotide probe and a target nucleic acid to which is hybridized a helper oligonucleotide.
Claim 23
Dependent Claim
23 words
A hybrid nucleic acid comprising a hybrid of a nucleotide probe and a target nucleic acid to which is hybridized a helper oligonucleotide.
Claim 24
Dependent Claim
17 words
A hybrid according to claim 21 wherein the duplex is comprised of more than one helper oligonucleotide.
Claim 25
Dependent Claim
29 words
A kit for the detection of a target nucleic acid comprising a labeled nucleotide probe complementary to a nucleotide sequence in the target nucleic acid and a helper oligonucleotide.
Claim 26
Dependent Claim
29 words
A kit for the detection of a target nucleic acid comprising a labeled nucleotide probe complementary to a nucleotide sequence in the target nucleic acid and a helper oligonucleotide.
Claim 27
Dependent Claim
12 words
A kit according to claim 23 comprising two or more helper oligonucleotides.
Claim 28
Dependent Claim
20 words
A kit according to claim 24 wherein the kit contains two or more probes and two or more helper oligonucleotides.
Claim 29
Dependent Claim
13 words
A kit according to claim 25 wherein each probe has a helper oligonucleotide.
Claim 30
Dependent Claim
17 words
A kit according to claim 26 wherein one or more probes has two or more helper oligonucleotides.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1994-07-08 | 1995-01-07 | 1995-07-09 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-07-08 | 1999-01-07 | 1999-07-09 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-07-08 | 2003-01-07 | 2003-07-09 | $7700.00 | Window Closed / Lapsed |