Gen-probe, Inc.

Applied Gene Technologies, Inc.

Bayer Corporation

The United States of America

Autogenomics, Inc.

Nanibhushan Dattagupta

Daniel L Kacian

C Nagaraja Sridhar

Aditya Ranjan Das

Yathi M Naidu

Whei-Kuo Wu

Jasmin R Patel

Sherrol Hoffa McDonough

Paul Douglas Stull

Thomas B Ryder

Diane L McAllister

Marc Spingola

David Bruce Lackey

Kathleen Ann Becherer

Philip W Hammond

Steven Joel Brown

Elizabeth R Billyard

Ta-Chien Tseng

Jurgen Kocher

Keiichi Kamisango

Contemplated systems and methods allow analysis of multiple and distinct patient samples using a labeling scheme that entirely avoids carry-over of a label or reagent to the analytic platform, typically an addressable solid phase. In preferred aspects, a hybridization portion, a fluorophore, and/or a quencher are removed by a 5'-3&#x2032;-exonuclease activity of a polymerase from a reporter oligonucleotide to so remove the oligonucleotide from the pool of molecules that bind to the solid phase and/or or to provide signal differentiation by removal of a fluorophore or quencher.

Concurrent analysis of multiple patient samples using solid phase addressable multiplex test with high signal-to-noise ratio

A method, composition and kit for synthesizing multiple copies of a target nucleic acid sequence autocatalytically under conditions of substantially constant temperature, ionic strength, and pH are provided in which multiple RNA copies of the target sequence autocatalytically generate additional copies using a mixture of blocked and unblocked primers and/or promoter-primers to initiate DNA and RNA synthesis, preferably with reduced non-specific product formation. The invention is useful for generating copies of a nucleic acid target sequence for purposes that include assays to quantitate specific nucleic acid sequences in clinical, environmental, forensic and similar samples, cloning and generating probes.

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Methods for amplifying target nucleic acid sequences using a nucleic acid polymerase lacking 5' exonuclease activity and a set of oligonucleotide primers. Preferably, a primer array is used. The primer array contains two sets of primers. One set contains at least two complementary primers. The other set contains at least two sense primers. Using the described methods amplification can be carried out under essentially constant environmental conditions without the requirement for exonuclease activity or restriction endonu- clease activity.

Isothermal strand displacement nucleic acid amplification

Methods for amplifying target nucleic acid sequences using a nucleic acid polymerase lacking 5' exonuclease activity and a set of oligonucleotide primers. Preferably, a primer array is used. The primer array contains two sets of primers. One set contains at least two complementary primers. The other set contains at least two sense primers. Using the described methods amplification can be carried out under essentially constant environmental conditions without the requirement for exonuclease activity or restriction endonuclease activity.

This invention relates generally to nucleic acid hybridization analysis. More specifically, oligonucleotide probes containing hairpin structures, or arrays of such oligonucleotide probes immobilized on a solid support, that are suitable for hybridization analysis are provided. Methods for nucleic acid hybridization analysis using the probes or array of immobilized probes are also provided.

Nucleic acid hairpin probes and uses thereof

The present invention relates to methods and compositions for analyzing nucleic acids. In particular, the invention provides for methods and combinations for analyzing nucleic acids in a plurality of samples using a plurality of detectably different signature labels and a probe that is hybridizable to each of the target nucleic acids. The invention also provides for a method for quantifying a nucleic acid by analyzing the amount of a label, e.g., a photoactivatable label, attached to the target nucleic acid.

Methods and compositions for analyzing nucleic acids

This invention relates generally to nucleic acid hybridization analysis. More specifically, a method for detecting a point mutation in a DNA strand is provided, which method uses, inter alia, a test nucleic acid strand complementary to a target DNA strand, said nucleic acid strand comprises a sufficient number of ribonucleotide residues that span the position of said point mutation to be detected to form a target DNA strand/test nucleic acid strand duplex and RNase H cleavage of said target DNA strand/test nucleic acid strand duplex. Kits and arrays for detecting a point mutation in a DNA strand comprising test nucleic acid strand comprising a sufficient number of ribonucleotide residues that span the position of said point mutation to be detected are also provided.

Methods and compositions for analyzing nucleotide sequence mismatches using RNase H

This invention relates generally to nucleic acid hybridization analysis. More specifically, an oligonucleotide probe for hybridization analysis is provided, which probe comprises a nucleotide sequence that forms a hairpin structure having a double stranded segment and a single stranded loop, wherein at least a portion of said nucleotide sequences located within said double stranded segment and a portion of said nucleotide sequence located within said single stranded loop collectively form a region that is complementary to a target nucleotide sequence to be hybridized with. Arrays comprising the hairpin probes immobilized on a solid support and methods for nucleic acid hybridization analysis using the probes or array of immobilized probes are also provided. Methods for transcribing and/or amplifying a probe DNA sequence using a hairpin probe are further provided.

The present invention describes compositions and methods for releasing nucleic acids from cells in a form that is suitable for labeling/capture, amplification, or detection in a single reagent addition step. The compositions include a lipid, membrane fluidizing compound, enzyme for degrading cell structure, metal chelators, or one or more nucleic acid probes or primers complementary to the nucleic acid to be detected. The compositions are non-denaturing and non-inhibitory of enzymes or proteins that are used in nucleic acid release, amplification, labeling or detection. The invention also provides kits for performing the above methods.

Sample processing to release nucleic acids for direct detection

This invention relates to methods of amplifying nucleic acids to minimize contamination by products of earlier amplification reactions. More particularly, it relates to methods of using nucleic acid labels that inhibit further amplification of the amplicon, and compositions that are useful to accomplish this task. In particular, the present invention relates to photoreactive complexes of a binding ligand, a binding enhancer and a label.

Method of labeling a nucleic acid amplicon with simultaneous contamination prevention

The present invention provides a cocktail of reagents for nucleic acid amplification that are stabilized by inclusion of a reversible inhibitor of undesirable reactions. Such cocktail of reagents eliminates the requirement for separate preparation and quality control of each reagent used in a reaction. Methods to prepare stabilized cocktails and to use stabilized cocktails also are included. The stabilized cocktail compositions also can include reagents to release nucleic acid from cells and to label the nucleic acid, allowing detection of nucleic acid in a sample with a single reagent addition step. The invention also provides kits for performing the above methods.

Stabilization of nucleic acid amplification cocktails

Methods for amplifying target nucleic acid sequences using a nucleic acid polymerase lacking 5&prime; exonuclease activity and a set of oligonucleotide primers. Preferably, a primer array is used. The primer array contains two sets of primers. One set contains at least two complementary primers. The other set contains at least two sense primers. Using the described methods amplification can be carried out under essentially constant environmental conditions without the requirement for exonuclease activity or restriction endonuclease activity.

Growing community of inventors

San Diego, CA, United States of America

Nanibhushan Dattagupta