Disclosed herein are compositions and methods that involve inserting connector protein channels of bacteriophage DNA packaging motors into copolymeric membranes via liposome-polymer fusion, which can be used as nanopore sensors for biomedical applications such as high throughput protein sequencing or cancer diagnosis. For example, disclosed are compositions comprising a copolymeric membrane into which a connector protein channel of a bacteriophage packaging motor has been inserted.
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.
A kit which has a first and second component parts, which are adapted for connection with each other. The first component part has a first array of electrical sensors, two substantially parallel lateral walls on the sides of the electrical connectors, two rails extending along the sides of the array, a front contact point and an overhang for receiving the second part. The second component part has a second array of electrical sensors for connection to the first array, front end configured to fit between the lateral walls of the first connector, and lateral sides having rail reliefs to fit the rails of the first connector. Connection of the first component part and the second component part forms a shoulder that aligns to locate the second array of electrical connectors in correct position for connection to the first array of electrical connectors.
The invention relates to improving the movement of a target polynucleotide with respect to a transmembrane pore when the movement is controlled by a polynucleotide binding protein. The invention also relates to improved transmembrane pores and polynucleotide binding proteins.
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
C07K 14/35 - Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Mycobacteriaceae (F)
Described herein, among other things, is a method of sequencing, comprising: concatenating a plurality of fragments of genomic DNA to produce concatenated DNA; sequencing the concatenated DNA to produce a plurality of sequence reads, wherein at least some of the sequence reads comprise: at least the sequence of the 3′ and/or 5′ ends of a fragment that corresponds to the locus of interest and sequence of one or both of the fragments that flank the fragment in the concatenated DNA; and grouping the sequence reads that corresponds to the locus of interest using, for each of the grouped sequence reads: the 3′ and/or 5′ end sequences; and/or the flanking sequence.
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.
The invention provides a method of detecting a target polynucleotide in a sample comprising: (a) contacting the sample with a guide polynucleotide that binds to a sequence in the target polynucleotide and a polynucleotide-guided effector protein, wherein the guide polynucleotide and polynucleotide-guided effector protein form a complex with any target polynucleotide present in the sample; (b) contacting the sample with a membrane comprising a transmembrane pore; (c) applying a potential to the membrane; and (d) monitoring for the presence or absence of an effect resulting from the interaction of the complex with the transmembrane pore to determine the presence or absence of the complex, thereby detecting the target polynucleotide in the sample.
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.
The invention relates to a new method of characterising two or more target polynucleotides using a pore. The method involves sequentially attaching to a first polynucleotide one or more subsequent polynucleotides to form a concatenated polynucleotide.
The invention relates to a new method of determining the presence, absence or characteristics of an analyte. The analyte is coupled to a membrane. The invention also relates to nucleic acid sequencing.
The invention relates to new methods for synthesising polynucleotide molecules according to a predefined nucleotide sequence. The invention also relates to methods for the assembly of synthetic polynucleotides following synthesis, as well as systems and kits for performing the synthesis and/or assembly methods.
A pump for use in low-profile applications comprises a barrel for holding fluid; and a piston that converts a rotational driving force into a longitudinal driving motion within the barrel. The pump provides space saving advantages by reducing the need for external equipment and mechanisms around the pump for providing actuation or moving the actuating mechanism.
A61M 5/315 - Pistons; Piston-rods; Guiding, blocking or restricting the movement of the rod; Appliances on the rod for facilitating dosing
A61B 17/88 - Methods or means for implanting or extracting internal fixation devices
F04B 13/00 - Pumps specially modified to deliver fixed or variable measured quantities
F04B 7/06 - Piston machines or pumps characterised by having positively-driven valving in which the valving is performed by pistons and cylinders coacting to open and close intake or outlet ports the pistons and cylinders being relatively reciprocated and rotated
F04B 9/02 - Piston machines or pumps characterised by the driving or driven means to or from their working members the means being mechanical
A61M 5/145 - Pressure infusion, e.g. using pumps using pressurised reservoirs, e.g. by means of pistons
F04B 17/03 - Pumps characterised by combination with, or adaptation to, specific driving engines or motors driven by electric motors
B29L 31/26 - Sealing devices, e.g. packaging for pistons or pipe joints
F04B 53/14 - Pistons, piston-rods or piston-rod connections
The invention relates to a new method of determining the presence, absence or characteristics of an analyte. The analyte is coupled to a membrane. The invention also relates to nucleic acid sequencing.
A target polynucleotide is expanded. In respect of each nucleotide in the target polynucleotide, the target polynucleotide comprises clock nucleotides and at least one signal nucleotide in a predetermined order. The clock nucleotides have a predetermined sequence common to each nucleotide in the target polynucleotide. The at least one signal nucleotide is characteristic of the identity of the respective nucleotide in the target polynucleotide. During translocation of the expanded polynucleotide through a nanopore, electrical measurements dependent on the polynucleotide within the pore are made, to derive an analysis signal. Clock signals derived from the clock nucleotides are identified. Relative to the positions of the identified clock signals, nucleotide signals derived from the least one signal nucleotide are derived to analyse the target polynucleotide. The predetermined sequence of the clock nucleotides comprises a restriction site for a restriction enzyme and at least one further nucleotide that extends the predetermined sequence.
An analysis instrument comprises plural modules connected together over a data network, each module comprising an analysis apparatus operable to perform biochemical analysis of a sample. Each module comprises a control unit that controls the operation of the analysis apparatus. The control units are addressable to select an arbitrary number of modules to operate as a cluster for performing a common biochemical analysis. The control units communicate over the data network, repeatedly during the performance of the common biochemical analysis, to determine the operation of the analysis apparatus of each module required to meet the global performance targets, on the basis of measures of performance derived from the output data produced by the modules. The arrangement of the instrument as modules interacting in this manner provides a scalable analysis instrument.
An analysis instrument comprises plural modules connected together over a data network, each module comprising an analysis apparatus operable to perform biochemical analysis of a sample. Each module comprises a control unit that controls the operation of the analysis apparatus. The control units are addressable to select an arbitrary number of modules to operate as a cluster for performing a common biochemical analysis. The control units communicate over the data network, repeatedly during the performance of the common biochemical analysis, to determine the operation of the analysis apparatus of each module required to meet the global performance targets, on the basis of measures of performance derived from the output data produced by the modules. The arrangement of the instrument as modules interacting in this manner provides a scalable analysis instrument.
The invention relates to constructs comprising a transmembrane protein pore subunit and a nucleic acid handling enzyme. The pore subunit is covalently attached to the enzyme such that both the subunit and enzyme retain their activity. The constructs can be used to generate transmembrane protein pores having a nucleic acid handling enzyme attached thereto. Such pores are particularly useful for sequencing nucleic acids. The enzyme handles the nucleic acid in such a way that the pore can detect its component nucleotides by stochastic sensing.
C07K 14/31 - Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Micrococcaceae (F) from Staphylococcus (G)
C12N 9/16 - Hydrolases (3.) acting on ester bonds (3.1)
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.
Described herein, among other things, is a method of sequencing, comprising: concatenating a plurality of fragments of genomic DNA to produce concatenated DNA; sequencing the concatenated DNA to produce a plurality of sequence reads, wherein at least some of the sequence reads comprise: at least the sequence of the 3′ and/or 5′ ends of a fragment that corresponds to the locus of interest and sequence of one or both of the fragments that flank the fragment in the concatenated DNA; and grouping the sequence reads that corresponds to the locus of interest using, for each of the grouped sequence reads: the 3′ and/or 5′ end sequences; and/or the flanking sequence.
The invention relates to modified Dda helicases which can be used to control the movement of polynucleotides and are particularly useful for sequencing polynucleotides.
The invention relates to mutant forms of the outer membrane-located lipoprotein CsgG, in particular, modifications at one or more of positions Tyr51; Asn55; and Phe56. The invention also relates to analyte detection and characterisation using said mutant CsgG.
Arrangements are disclosed for measuring small electrical currents with high sensitivity, for example in the context of sensing molecular entities, for example via interactions between the molecular entities and a membrane protein inserted in an amphiphilic membrane. In one arrangement there is provided a current sensing circuit (52) configured to integrate the current output by a sensor element (56) during each of a plurality of sensing frames (62). In each sensing frame (62) first and second analogue samples of the integral are taken during first and second time windows (71,72). A readout circuit (54) processes the first and second analogue samples to output a digital output signal representing the current output by the sensor element (56). The processing comprises analogue to digital conversion processing and output processing. The output processing is performed exclusively during periods outside of the first and second time windows.
Methods and apparatus for forming apertures in a solid state membrane using dielectric breakdown are provided. In one disclosed arrangement a plurality of apertures are formed. The membrane comprises a first surface area portion on one side of the membrane and a second surface area portion on the other side of the membrane. Each of a plurality of target regions comprises a recess or a fluidic passage opening out into the first or second surface area portion. The method comprises contacting all of the first surface area portion of the membrane with a first bath comprising ionic solution and all of the second surface area portion with a second bath comprising ionic solution. A voltage is applied across the membrane via first and second electrodes in respective contact with the first and second baths comprising ionic solutions to form an aperture at each of a plurality of the target regions in the membrane.
B01D 67/00 - Processes specially adapted for manufacturing semi-permeable membranes for separation processes or apparatus
B01D 69/02 - Semi-permeable membranes for separation processes or apparatus characterised by their form, structure or properties; Manufacturing processes specially adapted therefor characterised by their properties
C25F 7/00 - Constructional parts, or assemblies thereof, of cells for electrolytic removal of material from objects; Servicing or operating
G01N 33/487 - Physical analysis of biological material of liquid biological material
The invention relates to a new method of determining the presence, absence or one or more characteristics of multiple analytes. The invention concerns coupling a first analyte to a membrane containing a detector and investigating the first analyte using the detector. The invention also concerns coupling a second analyte to the membrane and investigating the second analyte. The first analyte is uncoupled from the membrane prior to investigating the second analyte. The invention also relates to polynucleotide sequencing.
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.
The invention relates to hetero-oligomeric pores derived from Msp. The invention also relates to polynucleotide characterisation using the hetero-oligomeric pores.
C07K 14/35 - Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Mycobacteriaceae (F)
A61K 39/04 - Mycobacterium, e.g. Mycobacterium tuberculosis
To form a layer separating two volumes of aqueous solution, there is used an apparatus comprising elements defining a chamber, the elements including a body of non-conductive material having formed therein at least one recess opening into the chamber, the recess containing an electrode. A pre-treatment coating of a hydrophobic fluid is applied to the body across the recess. Aqueous solution, having amphiphilic molecules added thereto, is flowed across the body to cover the recess so that aqueous solution is introduced into the recess from the chamber and a layer of the amphiphilic molecules forms across the recess separating a volume of aqueous solution introduced into the recess from the remaining volume of aqueous solution.
C07K 14/35 - Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Mycobacteriaceae (F)
An analysis instrument comprises plural modules connected together over a data network, each module comprising an analysis apparatus operable to perform biochemical analysis of a sample. Each module comprises a control unit that controls the operation of the analysis apparatus. The control units are addressable to select an arbitrary number of modules to operate as a cluster for performing a common biochemical analysis. The control units communicate over the data network, repeatedly during the performance of the common biochemical analysis, to determine the operation of the analysis apparatus of each module required to meet the global performance targets, on the basis of measures of performance derived from the output data produced by the modules. The arrangement of the instrument as modules interacting in this manner provides a scalable analysis instrument.
G01N 33/00 - Investigating or analysing materials by specific methods not covered by groups
G01N 33/48 - Biological material, e.g. blood, urine; Haemocytometers
B01L 3/00 - Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
C12Q 1/00 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
An analysis apparatus for performing biochemical analysis of a sample using nanopores comprises: a sensor device that that supports plural nanopores, reservoirs holding material for performing the analysis; a fluidics system; and plural containers for receiving respective samples, all arranged in a cartridge that is removably attachable to an electronics unit arranged to generate drive signals to perform signal processing circuit to generate output data representing the results of the analysis. The fluidics system supplies samples selectively from the containers to the sensor device using a rotary valve. In one valve, a stator defines a plurality of first ports arranged around the rotational axis and a collection chamber extending in around the axis of rotation in communication with a second port. A rotor provides a passage extending between the collection chamber and a position in communication with any one of the plurality of first ports. In another valve a stator defining a plurality of first ports in an annular surface facing the rotational axis, and a rotor is mounted inside a liner arranged between the annular surface of the stator and a facing annular surface of the rotor. The liner has a greater compliance than the rotor and stator to facilitate sealing.
F16K 11/085 - Multiple-way valves, e.g. mixing valves; Pipe fittings incorporating such valves; Arrangement of valves and flow lines specially adapted for mixing fluid with all movable sealing faces moving as one unit comprising only taps or cocks with cylindrical plug
G01N 33/48 - Biological material, e.g. blood, urine; Haemocytometers
G01N 33/92 - Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving lipids, e.g. cholesterol
F16K 11/074 - Multiple-way valves, e.g. mixing valves; Pipe fittings incorporating such valves; Arrangement of valves and flow lines specially adapted for mixing fluid with all movable sealing faces moving as one unit comprising only sliding valves with pivoted closure members with flat sealing faces
A detachable electrical device can be formed from a kit comprising a pair of component parts adapted for connection to each other, wherein the connected components of the device may be subsequently disconnected, comprising: an array of electrical connectors, each electrical connector comprising an electrically conductive liquid; and an array of electrodes; wherein the arrays can be brought into contact with each other so as to provide a plurality of electrical connections between the electrically conductive liquid of the array of electrical connectors and the electrodes of the array of electrodes, and wherein the electrical connections may be subsequently broken by detaching the electrically conductive liquid from the electrodes of the array.
A pump for use in low-profile applications comprises a barrel for holding fluid; and a piston that converts a rotational driving force into a longitudinal driving motion within the barrel. The pump provides space saving advantages by reducing the need for external equipment and mechanisms around the pump for providing actuation or moving the actuating mechanism.
A61B 17/88 - Methods or means for implanting or extracting internal fixation devices
F04B 13/00 - Pumps specially modified to deliver fixed or variable measured quantities
A61M 5/315 - Pistons; Piston-rods; Guiding, blocking or restricting the movement of the rod; Appliances on the rod for facilitating dosing
F04B 9/02 - Piston machines or pumps characterised by the driving or driven means to or from their working members the means being mechanical
F04B 7/06 - Piston machines or pumps characterised by having positively-driven valving in which the valving is performed by pistons and cylinders coacting to open and close intake or outlet ports the pistons and cylinders being relatively reciprocated and rotated
F04B 17/03 - Pumps characterised by combination with, or adaptation to, specific driving engines or motors driven by electric motors
F04B 53/14 - Pistons, piston-rods or piston-rod connections
A61M 5/145 - Pressure infusion, e.g. using pumps using pressurised reservoirs, e.g. by means of pistons
B29L 31/26 - Sealing devices, e.g. packaging for pistons or pipe joints
C07K 14/35 - Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Mycobacteriaceae (F)
The invention relates to a new method of characterizing 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.
The invention relates to a method for modifying a template double stranded polynucleotide, especially for characterisation using nanopore sequencing. The method produces from the template a plurality of modified double stranded polynucleotides. These modified polynucleotides can then be characterised.
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
C07H 21/00 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
C12Q 1/6806 - Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
Analysis Of A Polymer A biochemical analysis system analyses polymers by taking measurements of a polymer from a sensor element comprising a nanopore during translocation of the polymer through the nanopore. When a polymer has partially translocated, the series of measurements is analysed using reference data derived from a reference sequence to provide a measure of similarity. Responsive to the measure of similarity, the sensor element may be selectively operated to eject the polymer and thereby make the nanopore available to receive a further polymer. Where the biochemical analysis system comprises an array of sensor elements and is takes measurements from sensor elements selected in a multiplexed manner, responsive to the measure of similarity, the biochemical analysis system ceases taking measurements from the currently selected sensor element and to starts taking measurements from a newly selected sensor element.
The invention relates to a method for method of characterizing, such as sequencing, a target double stranded polynucleotide. The polynucleotide is coupled to a membrane using at least two adaptors with different strengths of coupling to the membrane.
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
C07H 21/00 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
The invention relates to an improved method for characterising a template polynucleotide. The method involves using a polymerase to prepare a modified polynucleotide which makes it easier to characterise than the template polynucleotide.
The invention relates to improving the movement of a target polynucleotide with respect to a transmembrane pore when the movement is controlled by a polynucleotide binding protein. The invention also relates to improved transmembrane pores and polynucleotide binding proteins.
C07K 14/35 - Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Mycobacteriaceae (F)
The invention relates to new methods of controlling the movement of polynucleotides through transmembrane pores. The invention also relates to new methods of characterizing target polynucleotides using helicases.
The invention relates to a new method of characterising a target ribonucleic acid (RNA) involving forming a complementary polynucleotide. The method uses a transmembrane pore.
C12Q 1/6883 - Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
The invention relates to a method for modifying a template double stranded polynucleotide, especially for characterization using nanopore sequencing. The method produces from the template a plurality of modified double stranded polynucleotides. These modified polynucleotides can then be characterized.
The invention relates to constructs comprising a transmembrane protein pore subunit and a nucleic acid handling enzyme. The pore subunit is covalently attached to the enzyme such that both the subunit and enzyme retain their activity. The constructs can be used to generate transmembrane protein pores having a nucleic acid handling enzyme attached thereto. Such pores are particularly useful for sequencing nucleic acids. The enzyme handles the nucleic acid in such a way that the pore can detect its component nucleotides by stochastic sensing.
C07K 14/31 - Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Micrococcaceae (F) from Staphylococcus (G)
C12N 9/16 - Hydrolases (3.) acting on ester bonds (3.1)
The invention relates to new methods of moving helicases past spacers on polynucleotides and controlling the loading of helicases on polynucleotides. The invention also relates to new methods of characterizing target polynucleotides using helicases.
A target polynucleotide is expanded. In respect of each nucleotide in the target polynucleotide, the target polynucleotide comprises clock nucleotides and at least one signal nucleotide in a predetermined order. The clock nucleotides have a predetermined sequence common to each nucleotide in the target polynucleotide. The at least one signal nucleotide is characteristic of the identity of the respective nucleotide in the target polynucleotide. During translocation of the expanded polynucleotide through a nanopore, electrical measurements dependent on the polynucleotide within the pore are made, to derive an analysis signal. Clock signals derived from the clock nucleotides are identified. Relative to the positions of the identified clock signals, nucleotide signals derived from the least one signal nucleotide are derived to analyze the target polynucleotide. The predetermined sequence of the clock nucleotides comprises a restriction site for a restriction enzyme and at least one further nucleotide that extends the predetermined sequence.
An analysis instrument comprises plural modules connected together over a data network, each module comprising an analysis apparatus operable to perform biochemical analysis of a sample. Each module comprises a control unit that controls the operation of the analysis apparatus. The control units are addressable to select an arbitrary number of modules to operate as a cluster for performing a common biochemical analysis. The control units communicate over the data network, repeatedly during the performance of the common biochemical analysis, to determine the operation of the analysis apparatus of each module required to meet the global performance targets, on the basis of measures of performance derived from the output data produced by the modules. The arrangement of the instrument as modules interacting in this manner provides a scalable analysis instrument.
The invention relates to a new method of determining in a sample the presence, absence or concentration of one or more target analytes, such as micro-ribonucleic acids (microRNAs or miRNAs). The invention may therefore relate to a multiplex assay for determining the presence or absence of each target analyte in a group of multiple analytess. The invention uses one or more probes comprising a quadruplex and transmembrane pores.
C40B 30/04 - Methods of screening libraries by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding
G01N 33/487 - Physical analysis of biological material of liquid biological material
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
To form a layer separating two volumes of aqueous solution, there is used an apparatus comprising elements defining a chamber, the elements including a body of non-conductive material having formed therein at least one recess opening into the chamber, the recess containing an electrode. A pre-treatment coating of a hydrophobic fluid is applied to the body across the recess. Aqueous solution, having amphiphilic molecules added thereto, is flowed across the body to cover the recess so that aqueous solution is introduced into the recess from the chamber and a layer of the amphiphilic molecules forms across the recess separating a volume of aqueous solution introduced into the recess from the remaining volume of aqueous solution.
A sensor system (1) for measuring an electrical signal across a lipid bilayer is formed by a cell (2) and an electrical reader unit (3) which are connectable together. The cell (2) is capable of supporting a lipid bilayer across an aperture (11) in a membrane (10) and has a construction which is cheap to manufacture. The reader unit (3) is a portable device which monitors an electrical signal generated in the connected cell (2) to allow analysis of that electrical signal. The sensor system (1) is intended for use outside of a laboratory setting.
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.
The invention relates to a method for modifying a template polynucleotide for characterization, especially for nanopore sequencing. The method produces a modified polynucleotide which is complementary to the template polynucleotide at some positions and which contains universal or abasic nucleotides at the other, and in some instances predicable, positions. The resulting modified polynucleotide can then be characterized.
C12P 19/34 - Polynucleotides, e.g. nucleic acids, oligoribonucleotides
C07H 21/00 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
C07H 21/04 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
The invention relates to methods using constructs comprising a helicase and an additional polynucleotide binding moiety. The helicase is attached to the polynucleotide binding moiety and the construct has the ability to control the movement of a polynucleotide. The constructs can be used to control the movement of polynucleotides and are particularly useful for sequencing polynucleotides.
The invention relates to a method of characterising a target polynucleotide using a single-stranded binding protein (SSB). The SSB is either an SSB comprising a carboxy-terminal (C-terminal) region which does not have a net negative charge or a modified SSB comprising one or more modifications in its C-terminal region which decreases the net negative charge of the C-terminal region.
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.
The invention relates to a new method of determining in a sample the presence or absence of one or more analyte members of a group of two or more analytes. The invention therefore relates to a multiplex assay for determining the presence or absence of each analyte in a group of multiple analytes. The assay uses aptamers and transmembrane pores.
C12N 15/115 - Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith
C07H 21/04 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
69.
Hairpin loop method for double strand polynucleotide sequencing using transmembrane pores
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.
The invention relates to a new method of characterizing a target polynucleotide. The method uses a pore and an XPD helicase. The helicase controls the movement of the target polynucleotide through the pore.
C12Q 1/00 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
C12Q 1/25 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving enzymes not classifiable in groups
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
The invention relates to constructs comprising a transmembrane protein pore subunit and a nucleic acid handling enzyme. The pore subunit is covalently attached to the enzyme such that both the subunit and enzyme retain their activity. The constructs can be used to generate transmembrane protein pores having a nucleic acid handling enzyme attached thereto. Such pores are particularly useful for sequencing nucleic acids. The enzyme handles the nucleic acid in such a way that the pore can detect its component nucleotides by stochastic sensing.
C07K 14/31 - Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Micrococcaceae (F) from Staphylococcus (G)
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
An apparatus for supporting an array of layers of amphiphilic molecules, the apparatus comprising: a body (11), formed in a surface of the body, an array of sensor wells (10) capable of supporting a layer of amphiphilic molecules (30) across the sensor wells, the sensor wells each containing an electrode (12) for connection to an electrical circuit, and formed in the surface of the body between the sensor wells, flow control wells capable of smoothing the flow of a fluid across the surface.
A piston head for a syringe pump comprises a barrier portion (11) for driving fluid through a syringe pump barrel (30), wherein a peripheral section (13,14) of the barrier portion (11) is shaped to seal against the syringe pump barrel (30); and a re-silent member (15) arranged to resist deformation of the shaped peripheral section (13,14) of the barrier portion (11).
F16J 15/328 - Manufacturing methods specially adapted for elastic sealings
F16J 15/3208 - Sealings between relatively-moving surfaces with elastic sealings, e.g. O-rings with at least one lip provided with tension elements, e.g. elastic rings
B29L 31/26 - Sealing devices, e.g. packaging for pistons or pipe joints
F04B 53/14 - Pistons, piston-rods or piston-rod connections
The invention relates to a new method of characterizing a target polynucleotide. The method uses a pore and a RecD helicase. The helicase controls the movement of the target polynucleotide through the pore.
A pump for use in low-profile applications comprises a barrel for holding fluid; and a piston that converts a rotational driving force into a longitudinal driving motion within the barrel. The pump provides space saving advantages by reducing the need for external equipment and mechanisms around the pump for providing actuation or moving the actuating mechanism.
A61B 17/88 - Methods or means for implanting or extracting internal fixation devices
F04B 53/14 - Pistons, piston-rods or piston-rod connections
A61M 5/315 - Pistons; Piston-rods; Guiding, blocking or restricting the movement of the rod; Appliances on the rod for facilitating dosing
F04B 9/02 - Piston machines or pumps characterised by the driving or driven means to or from their working members the means being mechanical
F04B 7/06 - Piston machines or pumps characterised by having positively-driven valving in which the valving is performed by pistons and cylinders coacting to open and close intake or outlet ports the pistons and cylinders being relatively reciprocated and rotated
F04B 17/03 - Pumps characterised by combination with, or adaptation to, specific driving engines or motors driven by electric motors
B29C 70/68 - Shaping composites, i.e. plastics material comprising reinforcements, fillers or preformed parts, e.g. inserts by incorporating or moulding on preformed parts, e.g. inserts or layers
A61M 5/145 - Pressure infusion, e.g. using pumps using pressurised reservoirs, e.g. by means of pistons
B29L 31/26 - Sealing devices, e.g. packaging for pistons or pipe joints
The invention relates to a new method of determining the presence, absence or characteristics of an analyte. The analyte is coupled to a membrane. The invention also relates to nucleic acid sequencing.
The invention relates to a new method of characterizing a target polynucleotide. The method uses a pore and a Hel308 helicase or amolecular motor which is capable of binding to the target polynucleotide at an internal nucleotide. The helicase or molecular motor controls the movement of the target polynucleotide through the pore.
A one-way valve, comprises a valve housing; a valve member provided within the valve housing, the valve member being operative to open and close the valve and comprising a diaphragm with a central orifice to allow fluid to pass from one side of the diaphragm to the other; a valve inlet provided on a first side of the diaphragm; and a valve outlet provided on a second side of the diaphragm; wherein the configuration of the diaphragm and the valve housing biases the diaphragm to seal the inlet at rest.
F16K 7/17 - Diaphragm cut-off apparatus, e.g. with a member deformed, but not moved bodily, to close the passage with flat, dished, or bowl-shaped diaphragm arranged to be deformed against a flat seat the diaphragm being actuated by fluid pressure
F16K 25/00 - VALVES; TAPS; COCKS; ACTUATING-FLOATS; DEVICES FOR VENTING OR AERATING - Details relating to contact between valve members and seats
F16K 15/14 - Check valves with flexible valve members
A61K 39/00 - Medicinal preparations containing antigens or antibodies
C07K 14/35 - Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Mycobacteriaceae (F)
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
An analysis apparatus for performing biochemical analysis of a sample using nanopores comprises: a sensor device that supports plural nanopores, reservoirs holding material for performing the analysis; a fluidics system; and plural containers for receiving respective samples, all arranged in a cartridge that is removably attachable to an electronics unit arranged to generate drive signals to perform signal processing circuit to generate output data representing the results of the analysis. The fluidics system supplies samples selectively from the containers to the sensor device using a rotary valve.
F16K 11/085 - Multiple-way valves, e.g. mixing valves; Pipe fittings incorporating such valves; Arrangement of valves and flow lines specially adapted for mixing fluid with all movable sealing faces moving as one unit comprising only taps or cocks with cylindrical plug
F16K 11/06 - Multiple-way valves, e.g. mixing valves; Pipe fittings incorporating such valves; Arrangement of valves and flow lines specially adapted for mixing fluid with all movable sealing faces moving as one unit comprising only sliding valves
G01N 1/26 - Devices for withdrawing samples in the gaseous state with provision for intake from several spaces
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
B01L 3/00 - Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
G01N 33/487 - Physical analysis of biological material of liquid biological material
G01N 30/00 - Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography
F16K 11/074 - Multiple-way valves, e.g. mixing valves; Pipe fittings incorporating such valves; Arrangement of valves and flow lines specially adapted for mixing fluid with all movable sealing faces moving as one unit comprising only sliding valves with pivoted closure members with flat sealing faces
G01N 33/92 - Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving lipids, e.g. cholesterol
An analysis instrument comprises plural modules connected together over a data network, each module comprising an analysis apparatus operable to perform biochemical analysis of a sample. Each module comprises a control unit that controls the operation of the analysis apparatus. The control units are addressable to select an arbitrary number of modules to operate as a cluster for performing a common biochemical analysis. The control units communicate over the data network, repeatedly during the performance of the common biochemical analysis, to determine the operation of the analysis apparatus of each module required to meet the global performance targets, on the basis of measures of performance derived from the output data produced by the modules. The arrangement of the instrument as modules interacting in this manner provides a scalable analysis instrument.
An apparatus for sensing of an interaction of a molecular entity with a membrane protein in a lipid bilayer comprises an array of sensor elements (21) arranged to output an electrical signal that is dependant on occurrences of the interaction. A detection circuit (3) comprised detection channels (30) capable of amplifying an electrical signal from a sensor element. More sensor elements (21) are provided than detection channels (30), and detection channels (30) are selectively connected to sensor elements (21) that have acceptable quality of performance in that a lipid bilayer is formed and that an acceptable number of membrane proteins are inserted, on the basis of the amplified electrical signals that are output from the detection channels. This improves the efficiency of utilization of the detection channels, due to inefficiency in the utilization of the sensor elements, resulting in a reduction in the cost of the apparatus and the ability to perform sensing using relatively small samples.
A method of fabricating a membrane having a tampered pore, a polymeric membrane having a tapered pore, and uses of such polymeric membrane are disclosed. The membrane includes apertures of increasing diameter which are aligned with each other to form the tapered pore.
B32B 3/24 - Layered products essentially comprising a layer with external or internal discontinuities or unevennesses, or a layer of non-planar form; Layered products essentially having particular features of form characterised by a discontinuous layer, i.e. apertured or formed of separate pieces of material characterised by an apertured layer, e.g. of expanded metal
The invention provides method of covalently coupling two or more moieties, the method comprising: (a) providing a first moiety having covalently attached thereto (i) at least one first linker comprising a first hybridizable region and (ii) at least one first group capable of forming a covalent bond; (b) providing a second moiety having covalently attached thereto (i) at least one second linker comprising a second hybridizable region capable of hybridizing to the first hybridizable region and (ii) at least a second group capable of forming a covalent bond with the first group; (c) contacting the first and second moieties under conditions that allow the first and second hybridizable regions to hybridize and link the moieties; and (d) exposing the linked moieties to conditions that allow the formation of a covalent bond between the first and second groups.
C07K 14/31 - Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Micrococcaceae (F) from Staphylococcus (G)
The invention relates to a mutant α-hemolysin (α-HL) pore which is useful for detecting one or more nucleotides by stochastic sensing. The pore is particularly useful for sequencing DNA or RNA. A molecular adaptor that allows detection of the nucleotide(s) is covalently attached to the pore. The pore is specifically modified to facilitate positioning of the adaptor and may be modified to facilitate covalent attachment.
A nanopore device is described wherein is provided a sample input (110), an input chamber (120), and first and second sample chambers (130, 140) connected to the input chambers (120) via first and second nanopores (135, 145).
A method of fabricating a membrane having a tapered pore, a polymeric membrane having a tapered pore, and uses of such polymeric membrane are disclosed. The membrane includes apertures of increasing diameter which are aligned with each other to form the tapered pore.
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