Fluorescent labels and their use in separations
First Claim
1. A method of separating nucleic acid components of a nucleic acid multicomponent mixture, wherein each of the different nucleic acid components or combinations of components of interest are labeled with different labels comprising different fluorescer pairs, wherein said labels are characterized by:
- (1) having a donor-acceptor fluorescent pair bonded to an oligonucleotide chain with efficient energy transfer from said donor to said acceptor;
(2) each of the donor members of the labels absorbs at substantially the same wavelength as all of the other donor members and each of the acceptor members of the labels emits at a different wavelength compared to any of the other acceptor members; and
each of the different labels has substantially the same mobility in said separating as a result of varying the spacer of said donor-acceptor pair along said oligonucleotide chain, with the proviso that one of said labels can have the same fluorescer as the donor and the acceptor;
said method comprising;
binding different labels to different nucleic acid components or combinations of components of said multi-component mixture;
separating said nucleic acid components into individual fractions; and
detecting each of said labeled nucleic acid components by irradiating with a common light source at an absorption wavelength of said donors and detecting the fluorescence of each of said labels.
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Accused Products
Abstract
Compositions are provided comprising sets of fluorescent labels carrying pairs of donor and acceptor dye molecules, designed for efficient excitation of the donors at a single wavelength and emission from the acceptor in each of the pairs at different wavelengths. The different molecules having different donor-acceptor pairs can be modified to have substantially the same mobility under separation conditions, by varying the distance between the donor and acceptor in a given pair. Particularly, the fluorescent compositions find use as labels in sequencing nucleic acids.
169 Citations
11 Claims
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1. A method of separating nucleic acid components of a nucleic acid multicomponent mixture, wherein each of the different nucleic acid components or combinations of components of interest are labeled with different labels comprising different fluorescer pairs, wherein said labels are characterized by:
- (1) having a donor-acceptor fluorescent pair bonded to an oligonucleotide chain with efficient energy transfer from said donor to said acceptor;
(2) each of the donor members of the labels absorbs at substantially the same wavelength as all of the other donor members and each of the acceptor members of the labels emits at a different wavelength compared to any of the other acceptor members; and
each of the different labels has substantially the same mobility in said separating as a result of varying the spacer of said donor-acceptor pair along said oligonucleotide chain, with the proviso that one of said labels can have the same fluorescer as the donor and the acceptor;said method comprising; binding different labels to different nucleic acid components or combinations of components of said multi-component mixture; separating said nucleic acid components into individual fractions; and detecting each of said labeled nucleic acid components by irradiating with a common light source at an absorption wavelength of said donors and detecting the fluorescence of each of said labels. - View Dependent Claims (2, 5)
- (1) having a donor-acceptor fluorescent pair bonded to an oligonucleotide chain with efficient energy transfer from said donor to said acceptor;
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6. A kit comprising at least two fluorescent compounds for use as labels, each of said labels characterized by:
(1) comprising an acceptor-donor fluorescent pair bonded to a nucleic acid backbone chain, where the donor efficiently transfers energy to said acceptor for fluorescence of said acceptor;
(2) each of the donor members of said labels absorbs at substantially the same wavelength all of the other donor members and each of the acceptor members of said labels emits at a different wavelength compared to any of the other acceptor members; and
(3) each of said labels has substantially the same mobility in said separation resulting from varying the spacing of said donor-acceptor pair along said nucleic acid backbone, with the proviso that one of said labels can have the same fluorescer as the donor and the acceptor.- View Dependent Claims (3, 4, 7, 8, 9, 10, 11)
Specification