Roche Molecular Systems, Inc.

Hoffmann-la Roche Inc.

Cetus Corporation

Robert M Watson

David H Gelfand

Randall K Saiki

Pamela M Holland

Edward L Sheldon, Iii

Henry Rapoport

Michael Robert Chapman

Kary B Mullis

Corey H Levenson

Mary E Fisher

Richard M Snead

Stephen Howard Williams

A process of detecting a target nucleic acid using labeled oligonucleotides uses the 5' to 3&#x2032; nuclease activity of a nucleic acid polymerase to cleave annealed labeled oligonucleotide from hybridized duplexes and release labeled oligonucleotide fragments for detection. This process is easily incorporated into a PCR amplification, assay.

Homogeneous assay system

A process of detecting a target nucleic acid using labeled oligonucleotides uses the 5' to 3&#x2032; nuclease activity of a nucleic acid polymerase to cleave annealed labeled oligonucleotide from hybridized duplexes and release labeled oligonucleotide fragments for detection. This process is easily incorporated into a PCR amplification assay.

Process for discrimination of nucleic acid sequence variants

A process of detecting a target nucleic acid using labeled oligonucleotides uses the 5&prime; to 3&prime; nuclease activity of a nucleic acid polymerase to cleave annealed labeled oligonucleotide from hybridized duplexes and release labeled oligonucleotide fragments for detection. This process is easily incorporated into a PCR amplification assay.

A process of detecting a target nucleic acid using labeled oligonucleotides uses the 5' to 3' nuclease activity of a nucleic acid polymerase to cleave annealed labeled oligonucleotide from hybridized duplexes and release labeled oligonucleotide fragments for detection. This process is easily incorporated into a PCR amplification assay.

Reaction mixtures for detection of target nucleic acids

The present invention is directed to methods for controlling the light emission of an oligonucleotide labeled with a light-emitting label that are useful in nucleic acid detection assays. A reaction that results in the cleavage of single-stranded oligonucleotide probes labeled with a light-emitting label is carried out in the presence of a DNA binding compound that interacts with the label to modify the light emission of the label. The methods utilize the change in light emission of the labeled probe that results from degradation of the probe. The methods are applicable in general to assays that utilize a reaction that results in cleavage of oligonucleotide probes, and, in particular, to homogeneous amplification/detection assays wherein hybridized probe is cleaved concomitant with palmer extension. A homogeneous amplification/detection assay is provided which allows the simultaneous detection of the accumulation of amplified target and the sequence-specific detection of the target sequence.

Methods for in-solution quenching of fluorescently labeled

A process of detecting a target nucleic acid using labeled oligonucleotides which uses the 5' to 3' nuclease activity of a nucleic acid polymerase to cleave annealed labeled oligonucleotide from hybridized duplexes and thus releasing labeled oligonucleotide fragments for detection. This process is easily incorporated into a PCR amplification assay.

Nucleic acid detection by the 5'-3'exonuclease activity of polymerases

The present invention is directed to a process of detecting a target nucleic acid using labeled oligonucleotides. This process uses the 5' to 3' nuclease activity of a nucleic acid polymerase to cleave annealed labeled oligonucleotide from hybridized duplexes and release labeled oligonucleotide fragments for detection. This process is easily incorporated into a PCR amplification assay.

Homogeneous assay system using the nuclease activity of a nucleic acid

Nucleic acids may be labeled by complexing the alkylating moiety of a labeling reagent into a single-stranded nucleic acid to form a complex and activating the complex to cause covalent bonding between the reagent and the nucleic acid. Preferably, the labeled nucleic acid is a single-stranded hybridization probe for detecting nucleic acid sequences capable of hybridizing with a hybridizing region of the nucleic acid. Also preferably the label moiety is non-radioactive. The labeling reagent is of the formula:

Process for labeling single-stranded nucleic acids and hybridizaiton

Nucleic acids may be labeled by intercalating the alkylating intercalation moiety of a labeling reagent into a partially double-stranded nucleic acid to form a complex and activating the complex to cause covalent bonding between the reagent and the nucleic acid. Preferably, the labeled nucleic acid is a hybridization probe for detecting nucleic acid sequences capable of hybridizing with a hybridizing region of the nucleic acid. Also preferably the label moiety is non-radioactive. The labeling reagent is of the formula:

Growing community of inventors

Berkeley, CA, United States of America

Robert M Watson