Electronic Biosciences, Inc.

Roche Sequencing Solutions, Inc.

Quamtum Magnetics, Inc.

Electronics Biosciences, Inc.

Geoffrey A Barrall

Eric N Ervin

Jason Komadina

Prithwish Pal

Pirooz Parvarandeh

Charlotte Yang

Marcin Rojek

J William Maney

John Joseph Watkins

Takeshi Harada

Licheng Niu

Andrew Dennis Hibbs

Eric Takeshi Harada

Lowell J Burnett

Alan Gael Sheldon

Daniel K Lathrop

Melissa Ann Poquette

George John Carman

Ryan Dunnam

Systems and methods for inserting a single pore into a membrane under faradaic conditions are described herein. A stepped or ramped voltage waveform can be applied across the membranes of the cells of an array, where the voltage waveform starts at first voltage and increases in magnitude over a period of time to a second voltage. The voltage waveform has a polarity that maintains a first species of a redox couple in its current oxidation state. The first voltage is selected to be low enough to reduce the risk of damaging the membrane, while the rate of voltage increase is selected to provide sufficient time for the pores to insert into the membranes. Once a pore is inserted into the membrane, the voltage across the membrane rapidly drops, thereby reducing the risk of damaging the membrane even if the applied voltage between the electrodes is further increased.

Faradaic systems and methods for self-limiting protein pore insertion in amembrane

A method of forming a plurality of lipid bilayers over an array of cells in a nanopore based sequencing chip is disclosed. Each of the cells comprises a well. A first salt buffer solution with a first osmolarity is flowed over a cell in the nanopore based sequencing chip to substantially fill a well in the cell with the first salt buffer solution. A lipid and solvent mixture is flowed over the cell to deposit a lipid membrane over the well that encloses the first salt buffer solution in the well. A second salt buffer solution with a second osmolarity is flowed above the well to reduce the thickness of the lipid membrane, wherein the second osmolarity is a lower osmolarity than the first osmolarity such that an osmotic imbalance is created between a first volume inside the well and a second volume outside the well.

Osmotic imbalance methods for bilayer formation

Techniques for replacing nanopores within a nanopore based sequencing chip are provided. A first electrolyte solution is added to the external reservoir of the sequencing chip, introducing an osmotic imbalance between the reservoir and the well chamber located on the opposite side of a lipid bilayer membrane. The osmotic imbalance causes the membrane to change shape, and a nanopore within the membrane to be ejected. A second electrolyte solution is then added to the external reservoir to provide replacement nanopores and to restore the membrane shape. The replacement nanopores can be inserted into the membrane, effectively replacing the initial pore without causing the destruction of the membrane.

Removing and reinserting protein nanopores in a membrane using osmotic imbalance

Systems and methods for inserting a single pore into a membrane are described herein. A stepped or ramped voltage waveform can be applied across the membranes of the cells of an array, where the voltage waveform starts at first voltage and increases in magnitude over a period of time to a second voltage. The first voltage is selected to be low enough to reduce the risk of damaging the membrane, while the rate of voltage increase is selected to provide sufficient time for the pores to insert into the membranes. Once a pore is inserted into the membrane, the voltage across the membrane rapidly drops, thereby reducing the risk of damaging the membrane even if the applied voltage between the electrodes is further increased.

Systems and methods for self-limiting protein pore insertion in a membrane

A method of analyzing a molecule is disclosed. A lipid bilayer is formed such that it divides a first reservoir characterized by a first reservoir osmolarity from a second reservoir characterized by a second reservoir osmolarity. An electrolyte solution is flowed to the first reservoir that tends to make a first change to a ratio of the first reservoir osmolarity to the second reservoir osmolarity. A voltage is applied across the lipid bilayer, wherein the lipid bilayer is inserted with a nanopore, and wherein a net transfer of ions between the first reservoir and the second reservoir tends to make a second change to the ratio of the first reservoir osmolarity to the second reservoir osmolarity, and wherein the first change to the ratio and the second change to the ratio tends to counter-balance each other.

Counteracting osmotic imbalance in a sequencing cell

Provided are device components, devices and methods characterized by a high contrast signal to noise ratio (CNR).

High contrast signal to noise ratio device components

Provided herein are alpha hemolysin polypeptides comprising modified amino acid sequences that can reduce the rate of translocation of a polymer. Also provided herein are apparatuses and devices comprising modified hemolysin polypeptides. Also provided herein are methods of using modified alpha hemolysin proteins for use in characterizing and/or sequencing a polymer or for use as molecular sensors.

Modified alpha hemolysin polypeptides and methods of use

Provided are site specific chemically modified nanopore devices and methods for manufacturing and using them. Site specific chemically modified nanopore devices can be used for analyte sensing and analysis, for example.

Site specific chemically modified nanopore devices

Disclosed here are methods useful for incorporating protein into lipid bilayers using voltage induced insertion. The methods presented herein can decrease time and costs associated with incorporation of proteins into naturally derived or artificially created lipid bilayers. A method for incorporating a protein capable of translocating a ligand also is disclosed herein.

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San Jose, CA, United States of America

Geoffrey A Barrall