US4962045A
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Time-resolved fluorimetric detection of lanthanide labeled nucleotides
Assignee: Applied Biosystems LLC
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Filing: 1988-05-02
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Granted: 1990-10-09
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Expires: 2008-05-02
Abstract
Abstract Nucleotides conjugated with a chelating agent and labeled with a lanthanide, especially terbium, are readily detected by time-resolved fluorimetry during and following gel electrophoresis. The labeled nucleotide conjugates are very stable, highly fluorescent with a long radiative lifetime and remain fluorescent during dilution and during gel electrophoresis and since an enhancement solution is not required for detection, the labeled nucleotide conjugates can be detected on-line, if desired.
Inventors
- 👤 Enrico G. Picozza
- 👤 Steven S. Saavedra
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 17:20
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/533; C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6816; C12Q1/6869; G01N2458/00; G01N2458/40; Y; Y10; Y10S; Y10S436/00; Y10S436/80; Y10T; Y10T436/00; Y10T436/13; Y10T436/14; Y10T436/142222; Y10T436/143333
Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
44 words
In a method of time-resolved fluorimetric detection of fluorescent labeled nucleotide monomers, oligomers or polymers separated in a gel electrophoresis system the improvement wherein the fluorescent labeled nucleotide is a lanthanide chelate labeled covalent nucleotide conjugate and no enhancement solution is required for detection.
Claim 2
Dependent Claim
44 words
In a method of time-resolved fluorimetric detection of fluorescent labeled nucleotide monomers, oligomers or polymers separated in a gel electrophoresis system the improvement wherein the fluorescent labeled nucleotide is a lanthanide chelate labeled covalent nucleotide conjugate and no enhancement solution is required for detection.
Claim 3
Dependent Claim
16 words
A method according to claim 1 wherein the gel electrophoresis system is a polyacrylamide gel system.
Claim 4
Dependent Claim
18 words
A method according to claim 1 wherein the detecting of labeled nucleotides is for DNA or RNA sequencing.
Claim 5
Dependent Claim
21 words
A method according to claim 1 wherein the detecting of labeled nucleotides is by on-line detection in the gel electrophoresis system.
Claim 6
Dependent Claim
81 words
A method according to claim 1 wherein the nucleotide conjugate has the formula: Nuc-N*-Y-Z wherein Nuc is a nucleotide monomer, oligomer or polymer, N* is an amine nitrogen either intrinsic to the nucleotide or extrinsinc and introduced as a label prior to conjugation, Y is a chelating group capable of chelating a lanthanide and Z is an energy transfer moiety, and the lanthanide chelated to said nucleotide conjugate is selected from the group consisting of terbium, samarium, europium, dysoprosium or neodymium.
Claim 7
Dependent Claim
16 words
A method according to claim 5 wherein the gel electrophoresis system is a polyacrylamide gel system.
Claim 8
Dependent Claim
18 words
A method according to claim 5 wherein the detecting of labeled nucleotides is for DNA or RNA sequencing.
Claim 9
Dependent Claim
21 words
A method according to claim 5 wherein the detecting of labeled nucleotides is by on-line detection in the gel electrophoresis system.
Claim 10
Dependent Claim
21 words
A method according to claim 5 wherein the nucleotide conjugate has the formula: ##STR2## and the lanthanide chelated thereto is terbium.
Claim 11
Dependent Claim
16 words
A method according to claim 9 wherein the gel electrophoresis system is a polyacrylamide gel system.
Claim 12
Dependent Claim
18 words
A method according to claim 9 wherein the detecting of labeled nucleotides is for DNA or RNA sequencing.
Claim 13
Dependent Claim
21 words
A method according to claim 9 wherein the detecting of labeled nucleotides is by on-line detecting in the gel electrophoresis system.
Claim 14
Dependent Claim
40 words
In a method of separating fluorescent labeled nucleotide monomers, oligomers or polymers in a gel electrophoresis system the improvement wherein the fluorescent labeled nucleotide is a lanthanide chelate labeled covalent nucleotide conjugate and no enhancement solution is required for detection.
Claim 15
Dependent Claim
40 words
In a method of separating fluorescent labeled nucleotide monomers, oligomers or polymers in a gel electrophoresis system the improvement wherein the fluorescent labeled nucleotide is a lanthanide chelate labeled covalent nucleotide conjugate and no enhancement solution is required for detection.
Claim 16
Dependent Claim
16 words
A method according to claim 13 wherein the gel electrophoresis system is a polyacrylamide gel system.
Claim 17
Dependent Claim
16 words
A method according to claim 13 wherein the nucleotides are separated for DNA or RNA sequencing.
Claim 18
Dependent Claim
81 words
A method according to claim 13 wherein the nucleotide conjugate has the formula: Nuc-N*-Y-Z wherein Nuc is a nucleotide monomer, oligomer or polymer, N* is an amine nitrogen either intrinsic to the nucleotide or extrinsinc and introduced as a label prior to conjugation, Y is a chelating group capable of chelating a lanthanide and Z is an energy transfer moiety, and the lanthanide chelated to said nucleotide conjugate is selected from the group consisting of terbium, samarium, europium, dysprosium or neodymium.
Claim 19
Dependent Claim
16 words
A method according to claim 16 wherein the gel electrophoresis system is a polyacrylamide gel system.
Claim 20
Dependent Claim
16 words
A method according to claim 16 wherein the nucleotides are separated for DNA or RNA sequencing.
Claim 21
Dependent Claim
21 words
A method according to claim 16 wherein the nucleotide conjugate has the formula: ##STR3## and the lanthanide chelated thereto is terbium.
Claim 22
Dependent Claim
16 words
A method according to claim 19 wherein the gel electrophoresis system is a polyacrylamide gel system.
Claim 23
Dependent Claim
16 words
A method according to claim 19 wherein the nucleotides are separated for DNA or RNA sequencing.
References Cited (Backward Citations)
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| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1993-10-08 | 1994-04-09 | 1994-10-09 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1997-10-08 | 1998-04-09 | 1998-10-09 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2001-10-08 | 2002-04-09 | 2002-10-09 | $7700.00 | Window Closed / Lapsed |