Roche Molecular Systems, Inc.

Hoffmann-la Roche Inc.

Cetus Corporation

F. Hoffmann-la Roche Ag

Weyerhaeuser

Cea/institut De Genomique—centre National De Genotypage

Weyerhaeuser Company Limited

Roche Diagnostics Gmbh

Susanne Stoffel

Frances C. Lawyer

Moshe Benziman

Rony Tal

Anna Lisa Fear

Hing Wong

Roger D. Calhoon

David H Gelfand

Thomas W Myers

Randall K Saiki

Frances C Lawyer

Keith Bauer

Fred Lawrence Reichert

Robert M Watson

Pamela M Holland

Amar Gupta

Veeraiah Bodepudi

Edward Soh Smith

Richard D Abramson

Stephen Gordon Will

Shawn Lee Suko

Henry A Erlich

Kary B Mullis

Ellen H Fiss

Glenn T Horn

I Lawrence Greenfield

Christopher Lim Sigua

Hing C Wong

Alice M Wang

Lisa Vivian Kalman

Michael A Innis

Nancy J Schoenbrunner

Arie Ben-Bassat

Ivo Glynne Gut

James H Meade

Roger D Calhoon

Dieter Heindl

Herbert Von Der Eltz

Uwe Kobold

Russell G Higuchi

Shing Chang

Shirley Y Kwok

John J Sninsky

Keith A Bauer

John Niemiec

Lawrence I Greenfield

Mary Ann Brow

Kenneth B Myambo

Florence Mauger

Olga Budker

Thomas Koch

Roderic Fuerst

Nancy Jeneane Schönbrunner

Carita Maria Elfstrom

Christoph Steinbeck

Amar P Gupta

Nancy J Schönbrunner

The present application provides polynucleotides comprising 5'-tails with sequence segments useful for the detection of target nucleic acid sequences, and methods for their use in detecting target nucleic acids. The polynucleotides are used to amplify a subsequence of a target nucleic acid in the presence of one or more ribonucleotides. The ribonucleotides are incorporated into amplification products at regular intervals complementary to the 5&#x2032;-tail sequence segments. Cleavage of amplification products at the bond immediately 3&#x2032; to incorporated ribonucleotides produces detectably distinct fragments indicative of the presence or absence of a target nucleic acid.

Ribonucleotide tag nucleic acid detection

Disclosed are mutant DNA polymerases having improved extension rates relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.

DNA polymerases and related methods

The invention provides a method of determining the nucleotide sequence of a target nucleic acid using a reversibly terminating nucleotide that is modified at the 2' position.

Sequencing and genotyping using reversibly terminating nucleotides

Disclosed are mutant DNA polymerases having improved extension rates relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. The mutant polymerases overcome the inhibitory effects by an intercalating dye. Therefore, the mutant polymerases are useful in a variety of disclosed methods in combination with an intercalating dye. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.

The present invention provides methods of extending primer nucleic acids and sequencing target nucleic acids. The methods include the use of 2'-terminator nucleotides to effect chain termination. In addition to related reaction mixtures and kits, the invention also provides computers and computer readable media.

2'-terminator nucleotide-related methods and systems

Mutant DNA polymerases with improved pyrophosphorolysis activated polymerization (PAP) ability

The invention provides compositions that comprise nucleotides and/or nucleosides having blocking groups at 2'-positions of sugar moieties. Methods of synthesizing these nucleic acids are also provided.

Synthesis and compositions of 2'-terminator nucleotides

2'-Terminator nucleotide-related methods and systems

The present invention is directed to an improved analysis procedure for the comparative sequencing of nucleic acids using multistage mass spectrometry. More precisely, the invention is directed to a method enabling the de-novo sequencing of nucleic acid molecules using multistage mass spectrometry.

De-novo sequencing of nucleic acids

The present invention provides reaction mixtures that include blocked oligonucleotides comprising 2'-terminator nucleotides. The blocked oligonucleotides are rendered extendible when the 2&#x2032;-terminator nucleotides are removed from the oligonucleotides, e.g., via pyrophosphorolysis. The reaction mixtures can be used in various nucleic acid polymerization and/or amplification assays, among many other applications. In addition to reaction mixtures, the invention also provides related methods and reaction mixtures.

2'-terminator related pyrophosphorolysis activated polymerization

Modified thermostable DNA polymerases having enhanced efficiency for incorporating unconventional nucleotides such as those labeled with fluorescein family dyes are advantageous in many in vitro DNA synthesis applications. Such enzymes are particularly useful for use in chain termination nucleic acid sequencing protocols, as are native forms of such enzymes. Genes encoding the modified enzymes and methods for their production and use offer cost and efficiency advantages for DNA sequencing.

Thermostable DNA polymerases incorporating nucleoside triphosphates labeled with fluorescent dyes

Growing community of inventors

Oakland, CA, United States of America

David H Gelfand