Pulsed-multiline excitation for color-blind fluorescence detection
First Claim
1. A method of identifying sample components comprising:
- (a) preparing a sample comprising sample components, a first dye and a second dye;
(b) placing the sample in the beam path of a first excitation line and a second excitation line;
(c) sequentially firing the first excitation line and the second excitation line;
(d) collecting fluorescence signals from the samples as a function of time; and
(e) sorting the fluorescence by each excitation line'"'"'s on-time window, wherein the sample components are identified.
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Abstract
The present invention provides a technology called Pulse-Multiline Excitation or PME. This technology provides a novel approach to fluorescence detection with application for high-throughput identification of informative SNPs, which could lead to more accurate diagnosis of inherited disease, better prognosis of risk susceptibilities, or identification of sporadic mutations. The PME technology has two main advantages that significantly increase fluorescence sensitivity: (1) optimal excitation of all fluorophores in the genomic assay and (2) “color-blind” detection, which collects considerably more light than standard wavelength resolved detection. This technology differs significantly from the current state-of-the-art DNA sequencing instrumentation, which features single source excitation and color dispersion for DNA sequence identification. Successful implementation of the PME technology will have broad application for routine usage in clinical diagnostics, forensics, and general sequencing methodologies and will have the capability, flexibility, and portability of targeted sequence variation assays for a large majority of the population.
38 Citations
23 Claims
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1. A method of identifying sample components comprising:
- (a) preparing a sample comprising sample components, a first dye and a second dye;
(b) placing the sample in the beam path of a first excitation line and a second excitation line;
(c) sequentially firing the first excitation line and the second excitation line;
(d) collecting fluorescence signals from the samples as a function of time; and
(e) sorting the fluorescence by each excitation line'"'"'s on-time window, wherein the sample components are identified. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22)
- (a) preparing a sample comprising sample components, a first dye and a second dye;
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23. A method of identifying sample components comprising:
- (a) preparing a sample comprising sample components, a first dye and a second dye;
(b) placing the sample in the beam path of a first excitation line and a second excitation line;
(c) sequentially firing the first excitation line and the second excitation line;
(d) collecting fluorescence signals from the samples as a function of time, wherein said step of collecting fluorescence signals comprises measuring fluorescence emission using a non-dispersive detector; and
(e) sorting the fluorescence by each excitation line'"'"'s on-time window, wherein the sample components are identified.
- (a) preparing a sample comprising sample components, a first dye and a second dye;
Specification