Oxford Nanopore Technologies Limited

Oxford University Innovation Limited

Isis Innovation Limited

Katholieke Universiteit Leuven

Sharp Life Science (eu) Limited

Andrew John Heron

James White

Clive Gavin Brown

James Anthony Clarke

Lakmal Nishantha Jayasinghe

Elizabeth Jayne Wallace

Mark Bruce

Rebecca Victoria Bowen

Ruth Moysey

Szabolcs Soeroes

Daniel John Turner

Gavin Harper

Graham Hall

Daniel Ryan Garalde

Jason Robert Hyde

Domenico Caprotti

Christopher Peter Youd

John Hagan Pryce Bayley

David Alves

John Milton

Richard George Hambley

Luke McNeill

David Jackson Stoddart

Jonathan Bankes Pugh

Joseph Hargreaves Lloyd

Christopher James Wright

Stuart William Reid

Neil Roger Wood

Matthew Holden

Paul Raymond Mackett

Pedro Miguel Ortiz Bahamon

Giovanni Maglia

Gabriel Villar

Richard Alexander Gutierrez

Benjamin James Hadwen

David Needham

Sally Anderson

Marion Louise Crawford

Daniel George Fordham

Ramiz Iqbal Nathani

John Hagen Pryce Bayley

Richard Muscat

Paul Richard Moody

James Edward Graham

Matthew Alexander Holden

Mihaela Misca

Lorenzo Franceschini

Tine Brouns

Antonino Puglisi

Mark Ian Wallace

Jessica Mary May Knott

Anthony Clarke

Jessica Mary May Johnson

Mike Jennison

Mark John Bruce

David Antoni Alves

Jayne Wallace

Melania Slawa Strycharska

Luke Mcneill

An array of membranes comprising amphipathic molecules is formed using an apparatus comprising a support defining an array of compartments. Volumes comprising polar medium are provided within respective compartments and a layer comprising apolar medium is provided extending across the openings with the volumes. Polar medium is flowed across the support to displace apolar medium and form a layer in contact with the volumes, forming membranes comprising amphipathic molecules at the interfaces. In one construction of the apparatus, the support that comprises partitions which comprise inner portions and outer portions. The inner portions define inner recesses without gaps therebetween that are capable of constraining the volumes comprising polar medium contained in neighbouring inner recesses from contacting each other. The outer portions extend outwardly from the inner portions and have gaps allowing the flow of an apolar medium across the substrate.

Formation of array of membranes and apparatus therefor

Droplet interfaces are formed between droplets in an electro-wetting device comprising an array of actuation electrodes. Actuation signals are applied to selected actuation electrodes to place the droplets into an energised state in which the shape of the droplets is modified compared to a shape of the droplets in a lower energy state and to bring the two droplets into proximity. The actuation signals are then changed to lower the energy of the droplets into the lower energy state so that the droplets relax into the gap and the two droplets contact each other thereby forming a droplet interface. The use of sensing electrodes in the device permit electrical current measurements across the droplet interface. The sensing electrodes can be used for either (i) applying a reference signal during droplet actuation or (ii) recording electrical current measurements.

Droplet interfaces in electro-wetting devices

The invention relates to a new method of characterising a target polynucleotide. The method uses a pore and a Dda helicase. The helicase controls the movement of the target polynucleotide through the pore. The invention also relates to modified Dda helicases which can be used to control the movement of polynucleotides and are particularly useful for sequencing polynucleotides.

Modified helicases

The invention relates to mutant forms of lysenin. The invention also relates to analyte characterisation using lysenin.

Mutant lysenin pores

The invention relates to a sensing system comprising an electrowetting device, which electrowetting device comprises an array of actuation electrodes, and a control system configures to perform droplet operations on a system of droplets present in the sensing system. The invention also relates to a method of operating the sensing system of the invention. The invention also provides novel droplet constructs which can be made and manipulated in the sensing system of the invention.

Sensing system and method of operation

The invention relates to modified helicases with reduced unbinding from polynucleotides. The helicases can be used to control the movement of polynucleotides and are particularly useful for sequencing polynucleotides.

A sequence of polymer units in a polymer (), eg. DNA, is estimated from at least one series of measurements related to the polymer, eg. ion current as a function of translocation through a nanopore (), wherein the value of each measurement is dependent on a k-mer being a group of k polymer units (). A probabilistic model, especially a hidden Markov model (HMM), is provided, comprising, for a set of possible k-mers: transition weightings representing the chances of transitions from origin k-mers to destination k-mers; and emission weightings in respect of each k-mer that represent the chances of observing given values of measurements for that k-mer. The series of measurements is analysed using an analytical technique, eg. Viterbi decoding, that refers to the model and estimates at least one estimated sequence of polymer units in the polymer based on the likelihood predicted by the model of the series of measurements being produced by sequences of polymer units. In a further embodiment, different voltages are applied across the nanopore during translocation in order to improve the resolution of polymer units.

Method and system of estimating a sequence of polymer units

The invention relates to a new method of characterising a target polynucleotide using a pore. The method involves controlling the formation of secondary structure by the target polynucleotide after the polynucleotide has moved through the pore.

Methods of characterising target polynucleotides

The invention relates to new in vitro methods for synthesising a polymer, particularly a polynucleotide molecule, having a pre-defined sequence of units such as nucleotides. For synthesising a polynucleotide molecule the methods involve a process of extending a polynucleotide synthesis molecule with a transfer nucleotide. The methods additionally involve repeating the extension process multiple times to iteratively extend the polynucleotide molecule with multiple transfer nucleotides to generate a new polynucleotide molecule having a pre-defined nucleotide sequence. The invention also relates to in vitro methods of joining multiple synthetic polynucleotides following synthesis to form larger synthetic polynucleotides, as well as devices and systems for performing the extension, synthesis and assembly methods of the invention.

Polynucleotide synthesis method, kit and system

The invention relates to a method for sequencing a heteropolymeric target nucleic acid sequence that involves stochastic sensing. The invention also relates to a method for improving a pore for sequencing a target nucleic acid sequence by modifying one or more sites in the pore.

Method for sequencing a heteropolymeric target nucleic acid sequence

The invention relates to a new method of sequencing a double stranded target polynucleotide. The two strands of the double stranded target polynucleotide are linked by a bridging moiety. The two strands of the target polynucleotide are separated using a polynucleotide binding protein and the target polynucleotide is sequenced using a transmembrane pore.

Hairpin loop method for double strand polynucleotide sequencing using transmembrane pores

The invention relates to a new method of characterising a target RNA polynucleotide by taking one or more measurements as the target RNA polynucleotide moves with respect to a transmembrane pore. The movement is controlled by a DNA helicase. The invention also relates to a modified RNA construct wherein the RNA polynucleotide has been modified to increase DNA helicase binding thereto.

Method for nanopore RNA characterization

Apparatus and methods for controlling the insertion of a membrane channel into a membrane are disclosed. In one arrangement a first bath holds a first liquid in contact with a first surface of a membrane. A second bath holds a second liquid in contact with a second surface of the membrane. The membrane separates the first and second liquids. A first electrode contacts the first liquid. A second electrode contacts the second liquid. A driving unit applies a potential difference across the membrane via the first and second electrodes to promote insertion of a membrane channel into the membrane from the first liquid or the second liquid. A membrane voltage reduction unit is connected in series with the membrane. The driving unit applies a driving voltage across the membrane voltage reduction unit and the membrane, the driving voltage providing the potential difference across the membrane. The membrane voltage reduction unit is configured such that a reduction in resistance through the membrane caused by insertion of a membrane channel intrinsically increases a potential difference across the membrane voltage reduction unit thereby lowering the potential difference across the membrane. The lowering of the potential difference across the membrane is sufficient to prevent or reduce promotion of insertion of a further membrane channel.

Apparatus and methods for controlling insertion of a membrane channel into a membrane

The invention provides a droplet encapsulate comprising: a drop of a hydrophobic medium; a peripheral layer of non-polymeric amphipathic molecules around the surface of the drop; and an aqueous droplet within the peripheral layer, the aqueous droplet comprising: (a) an aqueous medium and (b) an outer layer of non-polymeric amphipathic molecules around the surface of the aqueous medium. The invention also provides processes for preparing the droplet encapsulates. Various uses of the droplet encapsulates are also described, including their use as drug delivery vehicles, in synthetic biology, and in the study of membrane proteins.

Multisomes: encapsulated droplet networks

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Oxford, United Kingdom

Andrew John Heron