Solution-based color compensation adjusted for temperature and electronic gains
First Claim
Patent Images
1. A method for determining the presence of at least two analytes in a sample, comprising:
- providing at least two detecting entities, each of said entities comprising a fluorescent label different from the fluorescent label of any of the other detecting entities, and each of said entities capable of specific binding with their respective analytes, contacting the sample with the detecting entities to allow specific binding of the analytes and detecting entities, exciting each of said labels with light having an appropriate wavelength for inducing fluorescence, determining values of fluorescence of the labels in at least two different spectral channels, and compensating said values for spectral overlap, wherein the compensating step includes correcting for temperature dependence of the fluorescent values.
4 Assignments
0 Petitions
Accused Products
Abstract
The present invention is directed to a method of analyzing at least two analytes at the same time using at least two fluorescent detecting entities. More particularly, the present invention relates to the use of fluorescently labeled hybridization probes to identify the genotypes of a nucleic acid sample at more than one locus by measuring fluorescence throughout a range of temperatures and correcting for temperature-dependent spectral overlap of the fluorescent probes.
-
Citations
18 Claims
-
1. A method for determining the presence of at least two analytes in a sample, comprising:
-
providing at least two detecting entities, each of said entities comprising a fluorescent label different from the fluorescent label of any of the other detecting entities, and each of said entities capable of specific binding with their respective analytes, contacting the sample with the detecting entities to allow specific binding of the analytes and detecting entities, exciting each of said labels with light having an appropriate wavelength for inducing fluorescence, determining values of fluorescence of the labels in at least two different spectral channels, and compensating said values for spectral overlap, wherein the compensating step includes correcting for temperature dependence of the fluorescent values. - View Dependent Claims (2, 3, 4, 5, 6)
-
-
7. A method of analyzing the genotype of multiple loci of nucleic acid sequences comprising the steps of:
-
providing a sample of nucleic acid having a first locus and a second locus, providing a first detecting entity, the first detecting entity comprising a first fluorescent label and having specificity for the first locus, providing a second detecting entity, the second detecting entity comprising a second fluorescent label and having specificity for the second locus, contacting the sample of nucleic acid with the first and second detecting entities, exciting the first and second entities with light having the appropriate wavelength to induce fluorescence of the first and second fluorescent labels, measuring throughout a range of temperatures a first fluorescent signal of the first detecting entity in a first spectral channel and a second fluorescent signal of the second detecting entity in a second spectral channel, and compensating the first and second signal values for temperature dependence of the fluorescent signals. - View Dependent Claims (8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
-
Specification