The Public Health Research Institute of the City of New York, Inc.

Columbia University

Rutgers, the State University of New Jersey

The Salk Institute for Biological Studies

Other

Phri Properties, Inc.

City University of New York

Gene-trak Systems

The Public Health Research Institute

Fred Russell Kramer

Sanjay Tyagi

Paul M Lizardi

Donald R Mills

Eleanor A Miele

Salvatore A Marras

Diana Vargas-Gold

David Alland

Leslie E Orgel

Barbara C Chu

Salvatore A E Marras

Cesar E Guerra

Alexander B Chetverin

Robert Vartikian

Paul Lizardi

Hilda M Buyoli

Vladimir D Axelrod

Ulf D Landegren

David Y Zhang

Jack W Szostak

Abraham Pinter

Gerald F Joyce

Ying Jie Wu

Karl A Drlica

Osama A Alsmadi

Amy Piatek

David Dubnau

Herman J Blok

Alexander B C{acute Over (h)}etverin

Hiyam Elhajj Trunfio

Oleg V Kurnasov

Lyubov A Ryabova

Igor Y Morosov

Elena V Volianik

Viktor I Ugarov

Jacqueline Vet

Alexander S Spirin

Salvatore AE Marras

This invention discloses multi-part primers for primer-dependent nucleic acid amplification methods. Also disclosed are multiplex assay methods, related reagent kits, and oligonucleotides for such methods.

Multiplex nucleic acid assay methods capable of detecting closely related alleles, and reagents therefor

This invention discloses multi-part primers for primer-dependent nucleic acid amplification methods. Also disclosed are primer-dependent nucleic acid amplification reactions, particularly DNA amplification reactions, reaction mixtures and reagent kits for such reactions. This invention relates to primer-dependent nucleic acid amplification reactions, particularly DNA amplification relations such as PCR, and primers, reaction mixtures and reagent kits for such reactions and assays employing same.

Highly selective nucleic acid amplification primers

This invention discloses multi-part primers for primer-dependent nucleic amplification methods. Also disclosed are primer-dependent nucleic acid amplification reactions, particularly DNA amplification reactions, reaction mixtures and reagent kits for such reactions. This invention relates to primer-dependent nucleic acid amplification reactions, particularly DNA amplification reactions such as PCR, and primers, reaction mixtures and reagent kits for such reactions and assays employing same.

This invention discloses multi-part primers for primer-dependent nucleic acid amplification methods. Also disclosed are primer-dependent nucleic acid amplification reactions, particularly DNA amplification reactions, reaction mixtures and reagent kits for such reactions. This invention relates to primer-dependent nucleic acid amplification reactions, particularly DNA amplification reactions such as PCR, and primers, reaction mixtures and reagent kits for such reactions and assays employing same.

The invention provides assays that can detect multiple genetic variants of a gene (e.g., a mycobacterium gene) in a sample using a pool (using 2, 3, 4, or more) of oligonucletide hybridization probes.

Assays for short sequence variants

Kits for highly multiplexed homogeneous in vitro screening assays for numerous possible nucleic acid targets, any of which might be present in a sample, that utilize fluorescent hybridization probes that are combinatorially coded from a panel of fluorophores by subdividing each probe into portions and differently labeling each portion such that, when portions are combined, each probe has a unique code. The kits may include reagents and primers for target amplification and real-time detection.

Homogeneous multiplex screening kits

A coding scheme for microcarriers suitable for use in distributed arrays includes labeling the carriers with quenched signaling hairpin molecules with any one of three to eight distinguishable fluorophores wherein the hairpins are of at least two types, most preferably three types, that open and fluoresce differentially as a chemical or physical condition, for example temperature, is changed. Mixtures of microcarriers having immobilized capture probes can be decoded by measuring fluorescence from said fluorophores under conditions under which only one type of hairpin is open, under which two types of hairpin are open, and so on. Mixtures of coded microcarriers with capture probes are used in assays for nucleic acids utilizing microarray methods.

Optically decodable microcarries, arrays and methods

The invention provides assays that can detect multiple genetic variants of a gene (e.g., a mycobacterial gene) in a sample using a pool (e.g., 2, 3, 4, or more) of oligonucleotide hybridization probes.

Highly multiplexed homogeneous in vitro screening assays for numerous possible nucleic acid targets, any of which might be present in a sample, utilize fluorescent hybridization probes that are combinatorially coded from a panel of fluorophores by subdividing each probe into portions and differently labeling each portion such that, when portions are combined, each probe has a unique code. The assays may include target amplification and real-time detection. Probe sets and kits containing additional assay reagents may be used to perform the screening assays.

Homogeneous multiplex screening assays and kits

Coalescence of cells or other membrane-bound entities is facilitated by anchoring an outwardly projecting first oligonucleotide in one member and an outwardly projecting second oligonucleotide, complementary to the first, in a second member and incubating under hybridizing conditions. Liposomes may be coalesced with cells to deliver hydrophilic agents thereto, such as DNA probes or drugs. Kits containing complementary oligonucleotides containing hydrophobic anchoring moieties may be used.

Oligonucleotide-facilitated coalescence

Non-competitive, quantitative amplification assay methods, including assays employing amplification by the polymerase chain reaction (PCR) process, for accurately measuring levels of target nucleic acid and sequences in samples and for ascertaining the relative amounts of cross-hybridizing alleles and mutants.

Non-competitive co-amplification methods

The present invention provides a replicatable and hybridizable recombinant single-stranded RNA probe molecule comprising: a recognition sequence for the binding of an RNA-directed RNA polymerase; a sequence required for the initiation of product strand synthesis by the polymerase; and a heteroloqus RNA sequence inserted at a specific site in the internal region of the recombinant molecule and complementary to an oligo or polynucleotide of interest. This invention also provides methods for determining the presence of concentration of an oligo- or polynucleotide of interest in a sample and for simultaneously determining the presence or concentration of several different oligo- or polynucleotides of interest in a sample.

Replicatable hybridizable recombinant RNA probes and methods of using same

For nucleic acid amplification including extension of primers by a DNA polymerase, high specificity primers are provided. The primers include a type of hairpin structure in which a single-stranded loop separates complementary 3&prime; and 5&prime; arms and in which the loop and the 3&prime; arm are complementary to the target nucleic acid. Amplification methods, assays and kits including such primers are included in the invention.

High specificity primers, amplification methods and kits

Ligation methods for manipulating nucleic acid stands and oligonucleotides utilizing hybridization arrays of immobilized oligonucleotides. The oligonucleotide arrays may be plain or sectioned, comprehensive or non-comprehensive. The immobilized oligonucleotides may in some cases be binary oligonucleotides having constant as well as variable segments. Some embodiments include amplification of ligated products.

Oligonucleotide arrays and their use for sorting, isolating, sequencing, and manipulating nucleic acids

Growing community of inventors

Riverdale, NY, United States of America

Fred Russell Kramer