Roche Sequencing Solutions, Inc.

Genia Technologies, Inc.

Other

Roche Molecular Systems, Inc.

Roche Diagnostics Gmbh

F. Hoffmann-la Roche Ag

Cynthia Ann Cech

Randall W Davis

Edward Shian Liu

Christos Tzitzilonis

Timothy Kellogg Craig

Andrew Trans

Eric Takeshi Harada

James Pollard

Liv Elisabeth Jensen

Anne Aguirre

Arkadiusz Bibillo

Aruna Ayer

Michael Dorwart

Peter Crisalli

Corissa Harris

Helen Franklin

Matthew Dipietro

Marshall Winston Porter

Dhruti Vasudev Dalal

Charlotte Yang

Tim Craig

Alexander Yang

Alexander Hyun-min Yang

Kirti Dhiman

Natalie B Chechelski Johnston

Joshua N Mabry

Hui Tian

John S Foster

Roger Chen

Robert A Yuan

Alexander Hyun-Min Yang

Liv Elizabeth Jensen

Timothy Kellog Cech

Timothy Kellog Craig

The present disclosure provides variant OmpG polypeptides, compositions comprising the OmpG variant polypeptides, and methods for using the variant OmpG polypeptides as nanopores for determining the sequence of single stranded nucleic acids. The variant OmpG nanopores reduce the ionic current noise versus the parental OmpG polypeptide from which they are derived and thereby enable sequencing of polynucleotides with single nucleotide resolution. The reduced ionic current noise also provides for the use of these OmpG nanopore variants in other single molecule sensing applications, e.g., protein sequencing.

OMPG variants

Disclosed are methods for isolating polymerase complexes from a mixture of polymerase complex components. The polymerase complexes can comprise a nanopore to provide isolated nanopore sequencing complexes. The methods relate to the positive and negative isolation of the polymerase complexes and/or nanopore sequencing complexes. Also disclosed is a nucleic acid adaptor for isolating active polymerase complexes, polymerase complexes comprising the nucleic acid adaptor, and methods for isolating active polymerase complexes using the nucleic acid adaptor.

Purification of polymerase complexes

OmpG variants

This disclosure provides a biochip comprising a plurality of wells. The biochip includes a membrane that is disposed in or adjacent to an individual well of the plurality of wells. The membrane comprises a nanopore, and the individual well comprises an electrode that detects a signal upon ionic flow through the pore in response to a species passing through or adjacent to the nanopore. The electrode can be a non-sacrificial electrode. A lipid bilayer can be formed over the plurality of wells using a bubble.

Methods for forming lipid bilayers on biochips

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

Alpha-hemolysin variants

A biochip for molecular detection and sensing is disclosed. The biochip includes a substrate. The biochip includes a plurality of discrete sites formed on the substrate having a density of greater than five hundred wells per square millimeter. Each discrete site includes sidewalls disposed on the substrate to form a well. Each discrete site includes an electrode disposed at the bottom of the well. In some embodiments, the wells are formed such that cross-talk between the wells is reduced. In some embodiments, the electrodes disposed at the bottom of the wells are organized into groups of electrodes, wherein each group of electrodes shares a common counter electrode. In some embodiments, the electrode disposed at the bottom of the well has a dedicated counter electrode. In some embodiments, surfaces of the sidewalls are silanized such that the surfaces facilitate the forming of a membrane in or adjacent to the well.

Process for biosensor well formation

Growing community of inventors

Newcastle, WA, United States of America

Cynthia Ann Cech