Patent US5853992A successfully refreshed from Google Patents.
US5853992A
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Cyanine dyes with high-absorbance cross section as donor chromophores in energy transfer labels
Assignee: University of California
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Filing: 1996-10-04
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Granted: 1998-12-29
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Expires: 2016-10-04
Abstract
Abstract Cyanine dyes are used as the donor fluorophore in energy transfer labels in which light energy is absorbed by a donor fluorophore and transferred to an acceptor fluorophore which responds to the transfer by emitting fluorescent light for detection. The cyanine dyes impart an unusually high sensitivity to the labels thereby improving their usefulness in a wide variety of biochemical procedures, particularly nucleic acid sequencing, nucleic acid fragment sizing, and related procedures.
Inventors
- 👤 Alexander N. Glazer
- 👤 Richard A. Mathies
- 👤 Su-Chun Hung
- 👤 Jingyue Ju
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 10:11
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6816; C12Q1/6818; C12Q1/6869
Claims Analysis
Total Claims: 28
Claim 1
Independent Claim
131 words
A fluorescent label comprising two fluorophores bonded to a backbone in energy transfer relationship whereby one of said fluorophores is a donor fluorophore and the other is an acceptor fluorophore, said donor fluorophore being a cyanine dye characterized by a quantum yield of between about 0.01 and about 0.25 and has the formula ##STR5## in which: X is O or S; Y is O or S; R 1 is a member selected from the group consisting of H, halo, and C 1 -C 6 alkyl; R 2 is a member selected from the group consisting of H, halo, and C 1 -C 6 alkyl; R 3 is C 1 -C 6 alkyl; and m is zero to 3; and said acceptor fluorophore having a quantum yield of at least about 0.3.
Claim 2
Dependent Claim
131 words
A fluorescent label comprising two fluorophores bonded to a backbone in energy transfer relationship whereby one of said fluorophores is a donor fluorophore and the other is an acceptor fluorophore, said donor fluorophore being a cyanine dye characterized by a quantum yield of between about 0.01 and about 0.25 and has the formula ##STR5## in which: X is O or S; Y is O or S; R 1 is a member selected from the group consisting of H, halo, and C 1 -C 6 alkyl; R 2 is a member selected from the group consisting of H, halo, and C 1 -C 6 alkyl; R 3 is C 1 -C 6 alkyl; and m is zero to 3; and said acceptor fluorophore having a quantum yield of at least about 0.3.
Claim 3
Dependent Claim
28 words
A fluorescent label in accordance with claim 1 in which said cyanine dye has an absorption maximum in the wavelength range of about 400 to about 900 nm.
Claim 4
Dependent Claim
47 words
A fluorescent label in accordance with claim 1 in which said cyanine dye has an absorption maximum in the wavelength range of about 400 to about 900 nm, and said acceptor fluorophore has an emission maximum in the wavelength range of about 450 to about 1,000 nm.
Claim 5
Dependent Claim
28 words
A fluorescent label in accordance with claim 1 in which said cyanine dye has an absorption maximum in the wavelength range of about 480 to about 550 nm.
Claim 6
Dependent Claim
47 words
A fluorescent label in accordance with claim 1 in which said cyanine dye has an absorption maximum in the wavelength range of about 480 to about 500 nm, and said acceptor fluorophore has an emission maximum in the wavelength range of about 500 to about 700 nm.
Claim 7
Dependent Claim
67 words
A fluorescent label in accordance with claim 1 in which: X is O; Y is O; R 1 is a member selected from the group consisting of H and C 1 -C 3 alkyl; R 2 is a member selected from the group consisting of H and C 1 -C 3 alkyl; R 3 is C 1 -C 3 alkyl; and m is 1, 2, or 3.
Claim 8
Dependent Claim
40 words
A fluorescent label in accordance with claim 1 in which: X is O; Y is O; R 1 is H or methyl; R 2 is H or methyl; R 3 is C 1 -C 3 alkyl; and m is 1.
Claim 9
Dependent Claim
36 words
A fluorescent label in accordance with claim 1 in which: X is O; Y is O; R 1 is H or methyl; R 2 is H or methyl; R 3 is ethyl; and m is 1.
Claim 10
Dependent Claim
32 words
A fluorescent label in accordance with claim 1 in which: X is O; Y is O; R 1 is methyl; R 2 is methyl; R 3 is ethyl; and m is 1.
Claim 11
Dependent Claim
33 words
A fluorescent label in accordance with claim 1 in which said backbone is a polymerized chain of monomeric units selected from the group consisting of purine and pyrimidine mononucleotides and hybridizing analogues thereof.
Claim 12
Dependent Claim
21 words
A fluorescent label in accordance with claim 1 in which said backbone is comprised of monomeric nucleoside units linked through N-(2-aminoethyl)glycine.
Claim 13
Dependent Claim
15 words
A fluorescent label in accordance with claim 1 in which said backbone is an oligonucleotide.
Claim 14
Dependent Claim
23 words
A fluorescent label in accordance with claim 12 in which said donor fluorophore is bonded to the 5' terminal unit of said oligonucleotide.
Claim 15
Dependent Claim
20 words
A fluorescent label in accordance with claim 12 in which said oligonucleotide is from 5 to 30 nucleotides in length.
Claim 16
Dependent Claim
20 words
A fluorescent label in accordance with claim 12 in which said oligonucleotide is from 5 to 20 nucleotides in length.
Claim 17
Dependent Claim
28 words
A fluorescent label in accordance with claim 12 in which said cyanine dye and said acceptor fluorophore are bonded to nucleotides separated by 1 to 29 intervening nucleotides.
Claim 18
Dependent Claim
28 words
A fluorescent label in accordance with claim 12 in which said cyanine dye and said acceptor fluorophore are bonded to nucleotides separated by 1 to 15 intervening nucleotides.
Claim 19
Dependent Claim
28 words
A fluorescent label in accordance with claim 12 in which said cyanine dye and said acceptor fluorophore are bonded to nucleotides separated by 4 to 12 intervening nucleotides.
Claim 20
Dependent Claim
17 words
A fluorescent label in accordance with claim 1 in which said acceptor fluorophore is a xanthene compound.
Claim 21
Dependent Claim
25 words
A fluorescent label in accordance with claim 1 in which said acceptor fluorophore is a member selected from the group consisting of fluoresceins and rhodamines.
Claim 22
Dependent Claim
33 words
A fluorescent label in accordance with claim 1 in which said cyanine dye is 3-(ε-carboxypentyl)-3'-ethyl-5,5'-dimethyloxacarbocyanine or 3-(ε-carboxypentyl)-3'-ethyl-oxacarbocyanine, and said acceptor fluorophore is a member selected from the group consisting of fluoresceins and rhodamines.
Claim 23
Dependent Claim
38 words
A fluorescent label in accordance with claim 1 in which said cyanine dye is 3-(ε-carboxypentyl)-3'-ethyl-5,5'-dimethyloxacarbocyanine or 3-(ε-carboxypentyl)-3'-ethyl-oxacarbocyanine, and said acceptor fluorophore is a member selected from the group consisting of FAM, R6G, TAMRA, ROX, JOE, and rhodamine 110.
Claim 24
Dependent Claim
39 words
A fluorescent label in accordance with claim 1 in which said cyanine dye is bonded to said backbone by a linking group having the formula ##STR6## in which n is 1 to 10, and p is 1 to 10.
Claim 25
Dependent Claim
21 words
A fluorescent label in accordance with claim 23 in which n is 1 to 6 and p is 2 to 8.
Claim 26
Dependent Claim
21 words
A fluorescent label in accordance with claim 23 in which n is 4 to 6 and p is 3 to 6.
Claim 27
Dependent Claim
17 words
A fluorescent label in accordance with claim 23 in which n is 5 and p is 6.
Claim 28
Dependent Claim
42 words
A fluorescent label in accordance with claim 1 in which said cyanine dye has the formula ##STR7## said backbone is an oligonucleotide, and said cyanine dye is bonded to a nucleotide of said backbone by a linking group having the formula ##STR8##
References Cited (Backward Citations)
US4996143A
Cited
US5188934A
Cited
US5326692A
Cited
US5326692B1
Cited
WO1994017397A1
Cited
WO1995021266A1
Cited
WO1996015270A1
Cited
EP0731178A2
Cited
GB2301833A
Cited
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| 3.5 Years (Year 4) | 2001-12-28 | 2002-06-29 | 2002-12-29 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2005-12-28 | 2006-06-29 | 2006-12-29 | $3760.00 | Window Closed / Lapsed |
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