Life Technologies Corporation

Hua Yu

Abraham Rosenbaum

Jeremy Gray

Collyn Seeger

Theo Nikiforov

The present disclosure provides methods, compositions and kits as well as systems for manipulating nucleic acids, including implementing isothermal amplification, such as recombinase-polymerase amplification (RPA), of a nucleic acid template using a pre-seeded solid support. Provided are rapid and efficient methods for generating template nucleic acid molecules comprising specific nucleotide sequence bound to solid support. Such methods can be used, for example, in manipulating nucleic acids in preparation for analysis methods that utilize monoclonal populations of nucleic acids.

Methods and compositions for manipulating nucleic acids

In some embodiments, the disclosure relates generally to methods, as well as related compositions and kits for recombinase-mediated nucleic acid amplification, such as recombinase-polymerase amplification (RPA), of a nucleic acid template using at least one blocked primer that contains a 5' domain, at least one nucleotide that is cleavable by an RNase H enzyme, a 3&#x2032; domain, wherein the primer is not extendable by a polymerase, and wherein the 3&#x2032; domain has a length of 7-100 nucleotides, for example 10-30 nucleotides. These methods and the use of a blocked primer reduce or eliminate non-specific amplification products, such as primer dimers, which are generated in RPA reactions.

Growing community of inventors

Guilford, CT, United States of America

Hua Yu