Labeled Oligonucleotide Probes Used for Nucleic Acid Sequence Analysis

摘要

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.

主权项

1. 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.