Cetus Corporation

Hoffmann-la Roche Inc.

Roche Molecular Systems, Inc.

Xytronyx, Inc.

Specialty Laboratories, Inc.

The Perkin-elmer Corporation

Kary B Mullis

Henry A Erlich

Randall K Saiki

Glenn T Horn

Corey H Levenson

David H Gelfand

Henry Rapoport

Shirley Y Kwok

John J Sninsky

Richard A Leath

Norman Arnheim

Edward L Sheldon, Iii

David H Mack

Stephen J Scharf

Larry Johnson

Joseph T Widunas

Louis M Mezei

Timothy J Wennberg

Robert M Watson

Henry A Ehrlich

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

Kits for amplifying and detecting nucleic acid sequences, including a

There is disclosed herein a machine for performing nucleic acid amplification under computer control. The machine utilizes any one of a number of heating and cooling systems under control of a host computer which directs the heating and cooling systems to heat and cool a reaction-chamber-containing heat exchanger at appropriate times in the process. The reaction chambers are pre-loaded with the nucleic acid(s) to be amplified, a thermostable enzyme to catalyze polymerization, specific oligonucleotide primers, and four different nucleotide triphosphates. Also disclosed is the process for the amplification chain reaction implemented by the machine, which utilizes a thermostable enzyme.

System for automated performance of the polymerase chain reaction

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

Photochemical systems for the direct visualization of exposure to ultraviolet radiation that effect visible color changes involving a process in which a photoacid is formed upon irradiation of a nitro-substituted aromatic aldehyde with ultraviolet light and wherein proton transfer to a dye causes the dye to undergo a visible color change. The system undergoes such color change to an extent directly proportional to the cumulative amount of ultraviolet incident thereupon.

Systems for the visualization of exposure to ultraviolet radiation

The presence or absence of a nucleic acid sequence associated with AIDS in a sample containing one or more nucleic acids and suspected of containing such sequence can be detected by amplifying the sequence using primers to form extension products as templates and detecting the amplified product if it is present. This may be accomplished by adding a labeled hybridization probe to the amplified product either free in solution or after immobilization on a solid support. Exemplary primers and probes for amplifying and detecting AIDS virus are provided.

Primes and probes for the amplification and detection of aids associated

Apparatus and method for performing automated amplification of nucleic

The present invention provides a method for rapidly obtaining substantially pure DNA from a biological sample containing cells. The method involves gently lysing the membranes of the cells to yield a lysate containing genomic DNA in a high molecular weight form. The lysate is moved through a porous filter to selectively trap the high molecular weight DNA on the filter. The DNA is released from the filter using an aqueous solution to form a solution containing substantially purified DNA, from which the DNA may analyzed or recovered.

Rapid purification of DNA

The present invention provides a method for rapidly obtaining substantially pure DNA from a biological sample containing cells. The method involves gently lysing the membranes of the cells to yield a lysate containing genomic DNA in a high molecular weight form. The lysate is moved through a porous filter to selectively trap the high molecular weight DNA on the filter. The DNA is released from the filter using an aqueous solution to form a solution containing substantially purified DNA, from which the DNA is recovered.

The presence or absence of a nucleic acid sequence associated with one or more related viruses in a sample containing one or more nucleic acids and suspected of containing such sequence can be detected by amplifying the sequence using primers to form extension products as templates and detecting the amplified product if it is present. This may be accomplished by adding a labeled hybridization probe to the amplified product either free in solution or after immobilization on a solid support. Preferably the virus constitutes AIDs viruses and hepadnaviruses.

Detection of viruses by amplification and hybridization

System for the visualization of exposure to ultraviolet radiation

A process for amplifying any target nucleic acid sequence contained in a nucleic acid or mixture thereof comprises treating separate complementary strands of the nucleic acid with a molar excess of two oligonucleotide primers and extending the primers with a thermostable enzyme to form complementary primer extension products which act as templates for synthesizing the desired nucleic acid sequence. The amplified sequence can be readily detected. 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.

Process for amplifying, detecting, and/or cloning nucleic acid sequences

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 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:

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La Jolla, CA, United States of America

Kary B Mullis