Back to results
US 2014/0178877 A1Published application
Labeled Oligonucleotide Probes Used for Nucleic Acid Sequence Analysis
AssigneeRoche Molecular Systems, Inc.
InventorsConcordio Anacleto, Teodorica Bugawan, Nancy Schoenbrunner
Life of this patent
Estimating the term…
Abstract
The present invention is directed to methods for generating a labeled oligonucleotide probe that contains a fluorescent rhodamine-derived dye for use in PCR reactions to detect a target nucleic acid.
Claims12
A method of making a labeled oligonucleotide for use as a hybridization probe in a PCR reaction comprising:
- (a) providing a fluorescent dye that is a rhodamine derivative wherein said fluorescent dye is attached with a bifunctional linker molecule;
- (b) binding a reactive group to the fluorescent dye via the bifunctional linker molecule for incorporating said fluorescent dye within the oligonucleotide;
- (c) incorporating the fluorescent dye between two internal nucleotides of the oligonucleotide, with the proviso that the fluorescent dye is not incorporated at the 5′ terminus of the oligonucleotide.
A method for detecting the presence or absence of a target nucleic acid or a target allele of a nucleic acid in a test sample, comprising:
- performing a PCR reaction with the use of a labeled probe oligonucleotide that hybridizes with the target nucleic acid, wherein the labeled probe oligonucleotide is characterized by having:
- a fluorescent dye that is a rhodamine derivative which is attached with a bifunctional linker molecule;
- and
- a reactive group bound to the bifunctional linker molecule for incorporating the fluorescent dye between two internal nucleotides of the oligonucleotide, with the proviso that that the fluorescent dye is not incorporated at the 5′ terminus of the oligonucleotide;
- detecting the signal from the fluorescent dye wherein the intensity of said signal represents the presence or absence of the target nucleic acid.
2 more elements
Dependencies are inferred from the claim text.