Roche Molecular Systems, Inc.

Cetus Corporation

Hoffmann-la Roche Inc.

Randall K Saiki

David H Gelfand

Henry A Erlich

Kary B Mullis

Glenn T Horn

Robert M Watson

Pamela M Holland

Amar Gupta

Alison Tsan

Igor Kozlov

Susanne Stoffel

Frances C Lawyer

Norman Arnheim

Stephen J Scharf

Shanavaz L Nasarabadi

The present invention describes methods for performing higher multiplexed real-time PCR for detection and quantitation of target nucleic acids using tagged hydrolysis probes.

Methods for performing multiplexed real-time PCR

The present invention describes methods for performing higher multiplexed PCR for detection and quantitation of target nucleic acids using tagged hydrolysis probes.

Methods for performing multiplexed PCR

The present invention describes methods for performing higher multiplexed real-time PCR for detection and quantitation of target nucleic acids using TAGS hydrolysis probes.

Methods for performing multiplexed real-time PCR in a self-contained nucleic acid analysis pouch

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

The present invention is directed to a process for amplifying any target nucleic acid sequence contained in a nucleic acid or mixture thereof using a thermostable enzyme. The process comprises treating separate complementary strands of the nucleic acid with a molar excess of two oligonucleotide primers, extending the primers with a thermostable enzyme to form complementary primer extension products which act as templates for synthesizing the desired nucleic acid sequence, and detecting the sequence so amplified. The steps of the reaction can be repeated as often as desired and involve temperature cycling to effect hybridization, promotion of activity of the enzyme, and denaturation of the hybrids formed.

Kits for amplifying and detecting nucleic acid sequences

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 purified thermostable nucleic acid polymerase is obtained that has unique characteristics. Preferably the nucleic acid polymerase is DNA polymerase isolated from a Thermus aquaticus species and has a molecular weight of about 86,000-95,000 daltons. The thermostable nucleic acid polymerase may be native or recombinant and may be used in a temperature-cycling chain reaction wherein at least one nucleic acid sequence is amplified in quantity from an existing sequence with the aid of selected primers and nucleotide triphosphates. The nucleic acid polymerase is preferably stored in a buffer containing non-ionic detergents that lends stability to the nucleic acid polymerase. A preferred buffer contains glycerol, polyoxyethylated sorbitan monolaurate, ethoxylated nonyl phenol and gelatin.

Stabilized thermostable nucleic acid polymerase compositions containing

Kits for amplifying and detecting nucleic acid sequences, including a

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

Single or multiple nucleotide variations in nucleic acid sequence can be detected in nucleic acids by a process whereby the sample suspected of containing the relevant nucleic acid is repeatedly treated with primers, nucleotide triphosphates, and an agent for polymerization of the triphosphates and then denatured, in a process which amplifies the sequence containing the nucleotide variation if it is present. In one embodiment, the sample is spotted on a membrane and treated with a labeled sequence-specific oligonucleotide probe. Hybridization of the probe to the sample is detected by the label on the probe.

Process for detecting specific nucleotide variations and genetic

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.

Growing community of inventors

Richmond, CA, United States of America

Randall K Saiki