General Electric Company

Ge Healthcare Bio-sciences Corp.

Global Life Sciences Solutions Operations UK Ltd

Anuradha Sekher

John Richard Nelson

Robert Scott Duthie

Carl Williams Fuller

Gregory Andrew Grossman

Gregory Andrew Grossmann

Shiv R Kumar

Anup Sood

Paul Mitsis

R Scott Duthie

Rohini Dhulipala

Gyanendra Kumar

John Nelson

Disclosed are methods and kits for endonuclease-assisted DNA amplification reaction using decontaminated primer solutions that are pre-treated with a nuclease. Nucleic acid amplification assays that employ nuclease-resistant, inosine-containing primers, endonuclease V enzymes to introduce a nick into a target DNA comprising at least one inosine, and a DNA polymerase to generate amplicons of a target DNA are also disclosed.

Endonuclase-assisted isothermal amplification using contamination-free reagents

Methods of amplification, purification and detection of nucleic acid sequences especially RNA are described. One aspect of the method involves the hybridisation and subsequent ligation of a nucleic acid structure to the nucleic acid sequence desired to be manipulated. The methods require that the nucleic acid structure comprises a double stranded region and a single stranded region. The single stranded region is complementary to the RNA sequence of interest. The double stranded region may also contain additional functionalities which are then used subsequently in the method.

Ligation-based RNA amplification

Provided herein are mutant endonuclease V enzymes that are capable of nicking an inosine-containing DNA sequence. Nucleic acid assays and agents that employ such mutant endonuclease V enzymes to introduce a nick into a target DNA including one or more inosine, and uses a DNA polymerase to generate amplicons of a target DNA are also described.

Mutant endonuclease V enzymes and applications thereof

Provided herein are nucleic acid synthesis methods and agents that employ an endonuclease for example, endonuclease V, to introduce a nick into a target DNA including one or more inosine, and uses a DNA polymerase to generate amplicons of the target DNA.

Isothermal DNA amplification

The present invention provides methods, formulation and kits for the synthesis of long nucleic acid fragments. An improved PCR method is provided for amplifying long DNA fragments. In particular, a single thermostable DNA polymerase is used for the rapid amplification of over 10 kb long DNA fragments. Also provided is a method for extending long complementary DNA strands using this single enzyme formulation.

Single enzyme system for fast, ultra long PCR

The present invention relates generally to the use of terminal-phosphate-labeled nucleotides having three or more phosphates as substrates for nucleic acid polymerases and their use in DNA amplification. The labels employed are chemiluminescent, fluorescent, electrochemical and chromogenic moieties as well as mass tags and include those that are directly detectable, detectable after enzyme activation or feed into other processes to generate a different signal. The signal generated from the attached dyes may also be used to quantify the amount of amplification. Further provided are stabilizers that enhance the stability of terminal-phosphate labeled nucleoside polyphosphates in aqueous solutions and are useful for reducing non-enzymatic hydrolysis of these nucleotides, hence decrease background.

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Belle Mead, NJ, United States of America

Anuradha Sekher