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.
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 characterising target polynucleotides using helicases.
An array of membranes comprising amphipathic moleculesis 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 provides a method of forming a membrane between a first volume of polar medium and a second volume of polar medium, which method comprises: (a) providing a first volume comprising polar medium and a second volume comprising polar medium which are separated from one another by an apolar medium, wherein at least one of said first and second volumes comprises a layer comprising amphipathic molecules, at the interface between the polar medium and the apolar medium, wherein each of the amphipathic molecules comprises a first outer hydrophilic group, a hydrophobic core group, and a second outer hydrophilic group, wherein each of the first and second outer hydrophilic groups is linked to the hydrophobic core group; and (b) causing the first and second volumes to come into contact with one another to form a membrane comprising said amphipathic molecules between the first and second volumes. The invention also provides a system comprising a membrane between a first volume of a polar medium; and a second volume of a polar medium, which membrane comprises the amphipathic molecules, and wherein the first volume of polar medium is within an apolar medium.
B01D 69/00 - Semi-permeable membranes for separation processes or apparatus characterised by their form, structure or propertiesManufacturing processes specially adapted therefor
B01J 13/10 - Complex coacervation, i.e. interaction of oppositely charged particles
C07K 14/35 - Peptides having more than 20 amino acidsGastrinsSomatostatinsMelanotropinsDerivatives thereof from bacteria from Mycobacteriaceae (F)
C07K 17/00 - Carrier-bound or immobilised peptidesPreparation thereof
C12M 1/34 - Measuring or testing with condition measuring or sensing means, e.g. colony counters
G01N 33/48 - Biological material, e.g. blood, urineHaemocytometers
5.
HELICASE CONSTRUCT AND ITS USE IN CHARACTERISING POLYNUCLEOTIDES
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.
C07K 14/435 - Peptides having more than 20 amino acidsGastrinsSomatostatinsMelanotropinsDerivatives thereof from animalsPeptides having more than 20 amino acidsGastrinsSomatostatinsMelanotropinsDerivatives thereof from humans
C12Q 1/00 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions
7.
A METHOD FOR DETERMINING THE PRESENCE OF AN ANALYTE USING AN APTAMER
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.
The invention relates to a new method of characterising a target polynucleotide. The method uses a pore and a RecD helicase. The helicase controls the movement of the target polynucleotide through the pore.
C12M 1/34 - Measuring or testing with condition measuring or sensing means, e.g. colony counters
C12M 1/40 - Apparatus specially designed for the use of free, immobilised, or carrier-bound enzymes, e.g. apparatus containing a fluidised bed of immobilised enzymes
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
C12Q 1/6809 - Methods for determination or identification of nucleic acids involving differential detection
A sequence of polymer units in a polymer (3), 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 (1), wherein the value of each measurement is dependent on a k-mer being a group of k polymer units (4). 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.
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 determining the presence,absence or characteristics of an analyte. The analyte is coupled to a membrane. The invention also relates to nucleic acid sequencing.
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 com-prises 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 el-ements (21) that have acceptable quality of performance in that a lipid bilayer is formed and that an acceptable number of mem-brane proteins are inserted, on the basis of the amplified electrical signals that are output from the detection channels. This im-proves the efficiency of utilisation of the detection channels, due to inefficiency in the utilisation of the sensor elements, resulting in a reduction in the cost of the apparatus and the ability to perform sensing using relatively small samples.
G01D 5/252 - Selecting one or more conductors or channels from a plurality of conductors or channels, e.g. by closing contacts a combination of conductors or channels
G01N 27/00 - Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
G01N 33/487 - Physical analysis of biological material of liquid biological material
14.
ADAPTORS FOR NUCLEIC ACID CONSTRUCTS IN TRANSMEMBRANE SEQUENCING
The invention relates to adaptors for sequencing nucleic acids. The adaptors may be used to generate single stranded constructs of nucleic acid for sequencing purposes. Such constructs may contain both strands from a double stranded deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) template. The invention also relates to the constructs generated using the adaptors, methods of making the adaptors and constructs, as well as methods of sequencing double stranded 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
C12N 15/66 - General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligationUse of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease
C12Q 1/6806 - Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
To form a layer (11) separating two volumes of aqueous solution, especially a biolayer lipid membrane (BLM) there is used an apparatus comprising elements defining a chamber (7), the elements including a body (2) of non-conductivematerial having formed therein at least one recess (5) opening into the chamber, the recess containing an electrode (21). 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.
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