Stratagene

Stratagene California

Hologic, Inc.

Other

Agilent Technologies, Inc.

The Scripps Research Institute

Medical Research Council

Catalyst Assets LLC

California Institute of Technology

Cytyc Corporation

Medical Research Institute

Stratagene Cloning Systems

Union Bank of California, N.a.

Stratgene

Joseph A Sorge

Richard A Lerner

Gregory Paul Winter

William D Huse

Holly Hogrefe

Lutz Riechmann

Rebecca Lynn Mullinax

Jay Milton Short

Michael H Wigler

Andrew Firmin

Anne M Whalen

Connie Jo Hansen

John T Welsh

Michael McClelland

William C Zimlich, Jr

Alan L Greener

Michael Borns

Peter Edward Vaillancourt

Alexander S Belyaev

Carsten-Peter Carstens

Gothami Padmabandu

Reinhold Mueller

Scott Robert Happe

Craig Monell

Dan Shoemaker

Kerstien A Padgett

Michael Phillip Weiner

John C Bauer

Keith V Sylvester

Bahram Arezi

Daniel Davis Shoemaker

Ernest Beutler

Henry Ji

Carsten Carstens

Richard Gibbs

Douglas Tyler McKenzie

Nick Roelofs

Lisa Joy Hexdall

Scott B Happe

Hooly Hogrefe

The present invention provides methods, kits and compositions for the detection of an analyte. In the methods of the invention, a binding molecule coupled to a first polymerase is incubated with a modified polynucleotide template to form a copy of the modified polynucleotide template without any modified nucleotides. The unmodified copy is detected in a second amplification/primer extension reaction using a second polymerase that is unable to amplify the modified polynucleotide template. Detection of the unmodified copy is indicative of the presence and/or amount of the analyte in the sample. In place of the binding molecule coupled to a first polymerase, a pair of analyte-specific probes can also be used. The first analyte specific probe comprises a first binding moiety and a first portion of a first polymerase and the second analyte specific probe comprises a second binding moiety and a second portion of the first polymerase. When the binding moieties are bound to the analyte the first and second portions of the polymerase interact to form a functional polymerase complex that is used to form a copy of the modified polynucleotide template without any modified nucleotides.

Analyte detection assays

The subject invention relates to compositions comprising an enzyme mixture which comprises a first enzyme and a second enzyme, where the first enzyme comprises a DNA polymerization activity and the second enzyme comprises an 5'-3&#x2032; exonuclease activity and a reduced DNA polymerization activity. The invention also relates to the above compositions in kit format and methods for high fidelity DNA synthesis using the subject compositions of the invention.

High fidelity DNA polymerase compositions and uses thereof

The invention features a novel isolated Family B DNA polymerase, apolymerase JDF-3, and mutant recombinant forms thereof. Mutant polymerases of the invention are deficient in 3' to 5&#x2032; exonuclease activity and/or exhibit reduced discrimination against non-conventional nucleotides relative to the wild-type form of the polymerase.

Compositions and methods utilizing DNA polymerases

The invention encompasses methods for enriching for and identifying a polymorphism within a nucleic acid sample either by separating a subset of a nucleic acid sample or by selectively replicating a subset of a nucleic acid sample such that the polymorphism is contained within a nucleic acid population with reduced complexity, and then identifying the polymorphism within the enriched nucleic acid sample. Methods also are disclosed for enriching for and identifying a polymorphism by contacting a nucleic acid sample that includes a subset of nucleic acid molecules having a sequence that binds to a sequence-specific binding activity with a molecule having a sequence-specific binding activity under conditions which permit specific binding, such that the subset of nucleic acid molecules bound to the activity is enriched for nucleic acid molecules having the sequence recognized by the sequence-specific binding activity, and detecting a polymorphism with respect to a reference sequence in the subset of nucleic acid molecules.

Methods of enriching for and identifying polymorphisms

The present invention relates to a method for isolating from the immunological gene repertoire a gene coding for a receptor having the ability to bind a preselected ligand. Receptors produced by the gene isolated by the method, particularly catalytic receptors, are also contemplated.

Method for producing polymers having a preselected activity

The present invention provides methods, nucleic acids, compositions, and kits for detecting microRNA (miRNA) in samples. The methods comprise designing mRNA-specific primers, adding a polyA tail to the miRNA, and using reverse transcription and amplification to detect the miRNA. The nucleic acids, compositions, and kits typically comprise some or all of the components necessary to practice the methods of the invention.

Methods compositions, and kits for detection of microRNA

The invention relates to the generation and characterization of Archaeal DNA polymerase mutants with deficient 3'-5&#x2032; exonuclease activity and reduced base analog detection activity. The invention further provides for Archaeal DNA polymerase mutants with deficient 3&#x2032;-5&#x2032; exonuclease activity and reduced base analog detection activity containing additional mutations that modulate other DNA polymerase activities including DNA polymerization or reverse transcriptase activity. The invention also discloses methods and applications of DNA polymerases with deficient 3&#x2032;-5&#x2032; exonuclease activity and reduced base analog detection activity.

DNA polymerase compositions for quantitative PCR and methods thereof

The subject invention relates to compositions comprising an enzyme mixture which comprises a first enzyme and a second enzyme, where the first enzyme comprises a DNA polymerization activity and the second enzyme comprises an 3'-5&#x2032; exonuclease activity and a reduced DNA polymerization activity. The invention also relates to the above compositions in kit format and methods for high fidelity DNA synthesis using the subject compositions of the invention.

High fidelity DNA polymerase compositions and uses therefor

Method for tapping the immunological repertoire

The present invention is directed to methods of generating a signal indicative of the presence of said target nucleic acid sequence in a sample, comprising, incubating a sample comprising a an amplified target nucleic acid and a nucleic acid polymerase which substantially lacks 5' to 3&#x2032; exonuclease activity, adding a thermostable fen nuclease consisting of 5&#x2032; to 3&#x2032; exonuclease and/or endonuclease activity so as to cleave a cleavage structure and generate a signal.

Methods of detecting an amplified nucleic acid

The invention relates to and methods for generating a signal indicative of the presence of a target nucleic acid in a sample. The compositions and methods include a reverse transcriptase, a nuclease, an upstream primer and downstream probe.

Methods for detection of a target nucleic acid by forming a cleavage structure using a reverse transcriptase

The invention relates to compositions and methods for generating a signal indicative of the presence of a target nucleic acid in a sample, where the compositions and methods include an RNA polymerase, a FEN nuclease, and a probe.

Methods for detection of a target nucleic acid by forming a cleavage structure using an RNA polymerase

The invention relates to compositions and methods of generating a signal indicative of the presence of a target nucleic acid in a sample, where the method includes forming a cleavage structure by incubating a sample comprising a target nucleic acid with a probe. The invention also includes the steps of cleaving the cleavage structure with a nuclease to release a nucleic acid fragment to generate a signal, wherein generation of the signal is indicative of the presence of a target nucleic acid in a sample.

Cleavage of nucleic acids

Growing community of inventors

Del Mar, CA, United States of America

Joseph A Sorge