Roche Sequencing Solutions, Inc.

Columbia University

Genia Technologies, Inc.

Nautilus Subsidiary, Inc.

Roche Diagnostics Gmbh

F. Hoffmann-La Roche Ag

Michael Dorwart

Daniel Korenblum

Cynthia Ann Cech

Christos Tzitzilonis

Timothy Kellogg Craig

Marshall Winston Porter

Liv Elisabeth Jensen

Alexander Hyun-min Yang

Jingyue Ju

Randall W Davis

Dmitriy Gremyachinskiy

Parag Mallick

Stephen P Hendricks

Roger Chen

Arkadiusz Bibillo

Sergey M Kalachikov

Torri Elise Rinker

Janine Mok

Tural Aksel

Elvis Ikwa

Gregory Kapp

Irina Morozova

Rachel Galimidi

Sonal Tonapi

Joshua Simon Klein

Markus Burns

Julia Robinson

Cassandra Stawicki

Roger Ja Chen

Alexander Hyun-Min Yang

Liv Elizabeth Jensen

Timothy Kellog Cech

Timothy Kellog Craig

Described herein are variants of alpha-hemolysin having at least one mutation selected from T12R, T12K, N17R, N17K or combinations of T12 and N17 mutations. The variants in some embodiments may further comprise H144A. The α-hemolysin variants have a decreased time to thread.

Alpha-hemolysin variants with altered characteristics

An affinity reagent, having: (a) a retaining component such as a structured nucleic acid particle; and (b) one or both of (i) one or more label components attached to the retaining component, and (ii) one or more binding components attached to the retaining component.

Affinity reagents having enhanced binding and detection characteristics

Provided are methods of nucleic acid sample preparation for analysis, including analysis using nanopore sequencing applications. In particular provided are methods that allow for the creation of circular and linear DNA samples that contain asymmetric ends to ligate different and desired adaptors to the end of the sample, methods to create single nucleic acid molecules that automatically form concatemers for higher sequencing throughput; methods to create multiple copies (temperature dependent, multi-primed, open-fold replication) of one original nucleic acid sample, or to create single nucleic acid molecules containing multiple copies of more than one original nucleic acid sample (concatemerized samples) for improved accuracy and throughput are described; and methods for improved modified base calling.

Nucleic acid sample preparation methods

This disclosure provides chips, systems and methods for sequencing a nucleic acid sample. Tagged nucleotides are provided into a reaction chamber comprising a nanopore in a membrane. An individual tagged nucleotide of the tagged nucleotides can contain a tag coupled to a nucleotide, which tag is detectable with the aid of the nanopore. Next, an individual tagged nucleotide of the tagged nucleotides can be incorporated into a growing strand complementary to a single stranded nucleic acid molecule derived from the nucleic acid sample. With the aid of the nanopore, a tag associated with the individual tagged nucleotide can be detected upon incorporation of the individual tagged nucleotide. The tag can be detected with the aid of the nanopore when the tag is released from the nucleotide.

Nucleic acid sequences using tags

Described herein are variants of alpha-hemolysin having at least one mutation selected from T12R, T12K, N17R, N17K or combinations of T12 and N17 mutations. The variants in some embodiments may further comprise H144A. The &#x3b1;-hemolysin variants have a decreased time to thread.

Described herein are engineered alpha-hemolysin subunits having mutated oligomerization domains for assembling into heptameric nanopores in lipid bilayers.

Alpha-hemolysin variants

&#x3b1;-hemolysin variants with altered characteristics

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Mountain View, CA, United States of America

Michael Dorwart