Agilent Technologies, Inc.

Other

Alexander S Belyaev

Joseph A Sorge

Craig Monell

Nicholas Michael Sampas

Alicia Scheffer-Wong

The present invention relates to transposase adapters and uses thereof, including uses in preparing DNA molecules, in vitro amplification, sequencing of nucleic acids, and screening of DNA libraries for sequences of interest as well as nucleic acid delivery.

Plurality of transposase adapters for DNA manipulations

The present invention provides a simple and rapid method for preparing purified transposase complexes that are highly suited for fragmenting DNA. The method includes forming transposase complexes with oligonucleotide adapters in cell lysate, then purifying the complexes from the other substance in the cell lysate. Purification is accomplished using a specific binding pair, in which one member of the pair is bound to an oligonucleotide adapter of the complex and the other member of the pair is bound to a solid substrate. The bound complexes can be immediately used in DNA fragmentation reactions to produce solid substrate-bound DNA fragments, which can be used for any number of purposes, including as templates for amplification and sequencing.

Immobilized transposase complexes for DNA fragmentation and tagging

The present disclosure provides methods for barcoding a plurality of DNA samples using a microarray of barcode-containing transposase complexes. In some embodiments, the DNA samples and transposase complexes are present in aqueous droplets on the surfaces of opposing substrates, which allows a single DNA sample droplet to be combined with a single transposase-complex droplet. The barcoded DNA in the combined droplets can be used for any number of purposes, including as templates for amplification and sequencing.

Methods for on-array fragmentation and barcoding of DNA samples

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 present invention provides new compositions for transposase-mediated fragmenting and tagging DNA targets. The invention relates to the surprising discovery that use of manganese ions (Mn) in transposase reactions improves the transposase reaction. It also relates to the surprising discovery that Mgions can be used in a transposase reaction with wild-type and/or engineered transposases at levels much higher than previously thought. The invention provides for the use of naturally-occurring transposases in in vitro reactions, as well as improved schemes for cleaving, tagging, and amplifying target DNA.

Methods and compositions for DNA fragmentation and tagging by transposases

The present invention provides methods, kits and compositions for the detection of an analyte. In the methods of the invention, a complex is formed between two or more analyte specific probes (ASP) and an analyte. The analyte specific probes each have a portion of a polymerase which interact to form a functional polymerase complex upon binding of the ASP to the analyte. The functional polymerase complex then generates a detectable signal which is indicative of the presence and/or amount of the analyte in the sample.

Binary signaling assay using a split-polymerase

A recombinant genome comprising polynucleotides encoding at least two additional molecular chaperones and a protein of interest, recombinant baculovirus vectors providing molecular chaperones and a method for producing a foreign protein using said genomes and vectors.

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San Diego, CA, United States of America

Alexander S Belyaev