726 results about "Nanopore" patented technology

Provided herein are methods for high-accuracy sequencing of polymeric analytes, such as peptides, using nanopores or nanogaps. A method of the present disclosure may comprise generating a modified amino acid from an amino acid of the peptide, translocating the modified amino acid through or adjacent to a nanopore, and identifying the modified amino acid.

Systems and methods for sequencing polynucleotides using nanopores are disclosed. In some embodiments, a polynucleotide including a single-stranded region and a double-stranded region, in which the single-stranded region is disposed through a nanopore. The polynucleotide can be moved relative to the nanopore by electric forces while one or more structural locks keep the polynucleotide close to the nanopore. A characteristic signal based on nanopore ionic current blockade and associated with the regions of the polynucleotide at or near the nanopore recognition zone is measured and used to infer the nucleobase sequence of the polynucleotide. In some examples, the double-stranded region is extended by a polymerase, and the polymerase is removed from the polynucleotide. In some examples, signals measured under different applied voltages provide nonredundant information regarding the polynucleotide sequence.

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 non-linear seepage experiment method for gas breaking through a cover layer based on dark field imaging. The method comprises the following steps: placing a microfluidic rock chip on an objective table of a dark field scattering microscope; the injection end of the microfluidic rock chip is connected with a fluid injection module, and the outlet end of the microfluidic rock chip is connected with a downstream outflow pipeline; injecting the formation water solution into the micro-fluidic rock chip until the micro-fluidic rock chip is filled with the formation water solution; opening a gas injection system, and adjusting the dark field scattering microscope until a clear image of a micro-fluidic rock chip pore structure and carbon dioxide displacement formation water is obtained; injecting carbon dioxide with different pressures into the micro-fluidic rock chip, and observing nonlinear seepage migration images of carbon dioxide and formation water in the micro-fluidic rock chip; when it is observed that carbon dioxide migrates to the outlet end of the rock chip, the breakthrough moment is obtained, and the upstream and downstream pressure difference value of the microfluidic rock chip is the breakthrough pressure. According to the method, the nonlinear seepage of the carbon dioxide displacement formation water in the nanopores can be accurately observed, and the breakthrough pressure can be determined.

The invention discloses a nanopore sensor based on insect olfactory receptor protein as well as a construction method and application thereof, and belongs to the field of biosensors. According to the invention, the insect olfactory receptor protein is used as a sensitive element to be integrated into a nanopore measurement system, so that the nanopore sensor is obtained. By measuring a nanopore current signal, the prepared sensor is applied to detection of target odorant molecules. According to the method, the recognition capability of the insect olfactory receptor protein on odorant molecules and the single molecule level detection capability of the nanopores are utilized, expensive large experimental instruments and complex experimental operation steps are not needed, and sensitive, rapid and direct detection on odorant substances can be realized.

The invention belongs to the technical field of biomedical engineering, and particularly relates to a single-base resolution laminated two-dimensional material nanopore structure, a DNA sequencing device and a DNA sequencing method. The two-dimensional material nanopore structure comprises an insulating layer and conducting layers arranged on the upper side and the lower side of the insulating layer, through nanopores are formed in the insulating layer and the conducting layers, and the pore diameter of the nanopores is 2-5 nm; the insulating layer and the conductive layer are made of two-dimensional materials. The conductive layers on the two sides serve as upper and lower electrodes, and a power supply is connected between the upper and lower electrodes to form a phase conductive path which is provided with a longitudinal current detection element. A plurality of groups of two-dimensional material nanopore structures are stacked, and meanwhile, a plurality of groups of base pair signals are obtained. According to the invention, the advantages of small thickness, high physical strength and the like of the two-dimensional material nanopore are kept by using a few layers of two-dimensional material films, the vibration of the films is reduced by increasing the number of layers and improving the mechanical strength, so that the signal-to-noise ratio is improved, and the spatial resolution of the nanopore sensor is improved by designing the upper electrode and the lower electrode.

The invention relates to a variation identification method of a nanopore single molecule sequencing current signal, which comprises the following steps of: dividing an original current signal generated by nanopore single molecule sequencing into continuous time periods according to a sequence that a basic group passes through a nanopore, calculating an average signal rhythm, constructing a standard time rhythm when a standard basic group sequence passes through the nanopore, and identifying the variation of the nanopore single molecule sequencing current signal. Comparing the offset difference between the actually measured average signal rhythm and the standard time rhythm, wherein a difference salient region is a potential variation region; current signals of front and back neighborhoods are extracted, a current signal disturbance difference value is quantized, and a region with the disturbance difference value having obvious direction asymmetry characteristics is a variation region to be confirmed; the range is expanded, current signals are extracted, disturbance intensity is calculated, and when the disturbance intensity presents a gradually-enhanced or gradually-weakened continuous change trend, it is judged that the variation area to be confirmed has physical rationality; and determining a theoretical current signal by using the corresponding non-variation sequence, comparing the theoretical current signal with an actually measured current signal, calculating a current difference, and determining that the current difference exceeds a normal difference range as a real variation region.

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.

The invention relates to a target molecule identification and quantification method based on a solid nanopore detection technology, electronic equipment and a computer readable storage medium. The method comprises the following steps: (1) acquiring current signal data generated by a target molecule through a nanopore sensor within a period of time; (2) carrying out filtering processing on the current signal data; (3) identifying the filtered data to obtain a target via hole event set; (4) filtering the target via hole event set, and extracting signal data of the filtered events to obtain a target molecule recognition event; and (5) calculating the average interval time of occurrence of the target molecule recognition event, and obtaining the occurrence frequency of the target molecule based on the average interval time, namely the relative concentration of the target molecule. According to the method, automatic identification and quantitative analysis are realized aiming at mass signal data generated by a solid-state nanopore detection technology, and the analysis efficiency and accuracy are improved by optimizing a data identification method.

The invention relates to the technical field of transmission membranes, in particular to a transmission membrane based on dithienylethene and a preparation method of the transmission membrane. The transmission film comprises a base film and light-operated molecules, the base film is coated with the light-operated molecules, and the light-operated molecules have light response groups. A detection result shows that after ultraviolet irradiation, the transmission film can quickly realize complete conversion from an open loop to a closed loop; after visible light irradiation is recovered, the transmission film can efficiently and reversibly recover to an open-loop state. In addition, the transport membrane has a high on-off ratio to five ions of sodium, potassium, lithium, magnesium and calcium. The transmission membrane based on dithienylethene, which is prepared by taking terminal amino-modified dithienylethene as a basic unit, not only has abundant nano-channels, but also is endowed with the characteristic of regulating and controlling the opening and closing of the nano-pore by a photoresponse functional group, and the closing and opening of an ion channel can be accurately regulated and controlled through the alternate irradiation of ultraviolet light and visible light, so that the transmission membrane has a good application prospect. And the controllability of light-operated ion transmission is finally realized.

The present application describes in part isolated and modified Enterococci toxin (Epx) polypeptides, immunogenic compositions comprising Exp polypeptides, nanopores formed by Epx polypeptides, apparatus comprising Epx nanopores, and methods of use thereof.

The present invention relates to modified Dda helicases which can be used to control the movement of analytes such as polynucleotides. The modified Dda helicases are used in analyte detection and characterisation. The present invention also relates to novel protein pores and their uses in analyte detection and characterisation. The invention particularly relates to an isolated pore complex formed by a CsgG-like pore and a modified CsgF peptide, or a homologue or mutant thereof, thereby incorporating an additional channel constriction or reader head in the nanopore.

A system for controlling a translocation speed of a molecule through a nanopore may include a fluid chamber containing a solution with a magnetic susceptibility that is different from the magnetic susceptibility of the molecule, a nanopore situated in the fluid chamber, and at least one magnetic component configured to create a magnetic field gradient within the solution to control the translocation speed of a molecule through the nanopore. A system for controlling a translocation speed of a molecule through a nanopore may include a nanopore at least one magnetic component situated to create a magnetic field that causes the molecule to experience a rotational torque as it passes through the nanopore.

The invention relates to a YaxAB nanopore, a system comprising the nanopore and application of the YaxAB nanopore. The YaxAB nanopore of the present invention has a funnel-shaped three-dimensional structure so that the change in current blocking according to the movement of an analyte captured inside the nanopore can be maximized, thereby enabling very sensitive measurement even of the structure and minute changes in physical and chemical properties (including dynamic changes and the like) of the analyte. And can cause very strong electroosmosis with high cation selectivity, such that various types of analytes can be captured and analyzed without being limited to the type of total charge. In addition, with respect to the nanopore of the present invention, the amino acid sequence of the monomer constituting the nanopore can be adjusted to improve protein purification yield and control an oligomerization mechanism, and thus a YaxAB nanopore having a pore size that cannot be formed by a wild-type monomer can be prepared. Furthermore, with respect to the nanopores of the present invention, YaxAB nanopores of various pore diameters can be secured using one-time purification, and nanopores of desired pore diameters can be selectively secured. Thus, limitations of conventional biological nanopores having fixed pore diameters can be overcome, so that for the YaxAB nanopore of the present invention, the size of the nanopore can be adjusted according to the size of an analyte. Further, the YaxAB nanopores of the present invention have a very wide analyte analysis range of dynamic detection range from small analytes to large analytes, which is caused by a funnel-shaped three-dimensional structure having a wide inlet and a narrow outlet. In addition, the YaxAB nanopore of the present invention has the largest nanopore structure among the hitherto biological nanopores, and a large-size analyte, which cannot be detected by conventional biological nanopores, can be detected and analyzed in a single molecule on the basis of a funnel structure having a large inlet. Furthermore, the present invention has established an optimized composition of a membrane in which YaxAB nanopores can be stably present, and by using a sensor implemented on the basis thereof, it is possible to qualitatively and quantitatively analyze various types of analytes having relatively various sizes and charges at a single molecule level with very high resolution and sensitivity.

A nanopore sensing device comprises a planar structure provided with plural fluidic passages extending between the first and second chambers. The planar structure supports nanopores in membranes across respective passages and sensor electrodes are arranged to sense a fluidic electrical potential in respective passages between the nanopores and the second chamber. The passages comprise planar fluidic resistor portions between the sensor electrode and the second chamber, the planar fluidic resistor portions extending in a planar direction of the planar structure and being configured to form a fluidic resistor.

A multi-modal nanopore system for biosensing and sequencing. In some embodiments, the system includes a first nanopore device, the first nanopore device including: a first chamber and a second chamber in a planar substrate; a nano-fluidic channel connecting the first chamber and the second chamber; a first electrode and a second electrode forming a nanopore between the first electrode and the second electrode; an optical coupling element configured to couple an electromagnetic beam with the nanopore; a first bias circuit for applying a bias to the first electrode and the second electrode, to form a transverse electric field in the nanopore; and a current-measuring circuit configured to measure a current between the first electrode and the second electrode.

The invention belongs to the technical field of methods for characterizing target polynucleotides, and provides application of a biological nanopore protein and a mutant thereof in detection. The biological nanopore protein mutant is obtained by mutating wild type PANX1 nanopore protein, the amino acid sequence of the wild type PANX1 nanopore protein is shown as SEQ ID NO: 1, and the mutation means that one or more amino acids of the wild type PANX1 nanopore protein are mutated into common amino acids except original amino acids, or carrying out truncation or truncation substitution on the C region of the wild type PANX1 nanopore protein to obtain the PANX1 nanopore protein mutant. The wild type PANX1 nanopore protein can be normally expressed after mutation, compared with the wild type PANX1 nanopore protein, the biological nanopore protein mutant is stable in property, the C region is truncated, so that the pore diameter of the contraction region of the protein mutant is enlarged, and the pore channel is prevented from being blocked.

This invention provides a process for sequencing single-stranded DNA by employing a nanopore and modified nucleotides.

The present invention relates to a method for sequencing small non-coding RNAs on a nanopore platform. In this method, sequence-specific structured adapters are used to specifically capture small non-coding RNAs in a single ligation step reaction, thus enabling direct RNA sequencing.

The invention provides a biological nanopore sensor and a preparation method thereof. The method for preparing the biological nanopore sensor comprises the following steps: a) preparing hydrophilic protein into lipidosome; and b) mixing the lipidosome with the membrane layer to promote the lipidosome to be inserted into the membrane layer to obtain the biological nanopore sensor, and the hydrophilic protein has a nanopore channel. The method can solve the problem that the hydrophilic protein is difficult to prepare into the biological nanopore sensor in the prior art, and is suitable for the field of nanopore sequencing.

The present disclosure relates to systems and methods for protein analysis, and more particularly to nanopore systems, devices, and methods for single molecule protein analysis and sequencing. There is provided a method of translocating a target protein through a nanopore included in a membrane separating a fluid chamber of a nanopore system into a cis-side and a trans-side, comprising: (a) allowing a protein transferase in solution to capture and form a complex with the target protein to be translocated; (b) contacting a transposase-target protein complex with the cis-side of the nanopore and allowing the target protein to translocate to the trans-side; wherein the nanopore system has an electroosmotic force (EOF) from cis to trans, which is generated from a large net ionic current with respect to the total ionic current from cis to trans, so that the target protein is captured in the nanopore, and there is a transposase at the top of the nanopore that controls translocation.

The invention relates to an anthrax virus protein nanopore buffer solution system and biomolecule detection application, and belongs to the field of nanopore electrochemistry. The invention aims to solve the problem that an anthrax virus protein nanopore is unstable in electrochemical detection, and provides an anthrax virus protein nanopore buffer solution system and application thereof. The buffer solution is composed of trihydroxymethyl aminomethane and a compound I composed of specific cations and anions, the compound I comprises the cations such as NH4 < + >, K < + >, Li < + > and Mg < 2 + > and the anions such as Cl <->, HCOO <-> and SO4 < 2->, and the stability of nanopores is achieved by optimizing the concentration and the pH value. The method is applied to high-sensitivity and high-selectivity detection of different charged small molecules. According to the technical scheme, the stability of the anthrax virus protein nanopore is remarkably improved, the application prospect of the anthrax virus protein nanopore in the field of single molecule detection is widened, and a new thought and method are provided for development of a nanopore sensing technology.

The invention discloses a preparation method of a PVA / chitin porous nanosheet layered film for salinity gradient power generation, and belongs to the technical field of bio-based material functionalization. The preparation method comprises the following operation steps: firstly, preparing a chitin porous nanosheet by utilizing an acid steam combined ultrasonic strategy; modifying the surface of the chitin porous nanosheet with rich positive charges through quaternization reaction, and performing suction filtration and drying to obtain a porous nanosheet layered film; and finally, circularly freezing, thawing and packaging the porous nanosheet layered film by using polyvinyl alcohol hydrogel to prepare the PVA / chitin porous nanosheet layered film. According to the PVA / chitin porous nanosheet layered film, by means of combination of chitin nanosheet layer gap channels and micro-nano pore channels in nanosheet surfaces, ion transmission paths are increased, ion transmission resistance is reduced, the microstructure of the layered film is effectively stabilized jointly by means of the interlayer weak force effect and the hydrogen bond bonding effect between polyvinyl alcohol molecules, and therefore the film has the advantages of being capable of being used for preparing the composite film. And the optimal salinity gradient power generation output power density reaches 7.33 W / m < 2 >.

In one aspect, the disclosed technology relates to nanopore sequencing with a polynucleotide comprising a plurality of nucleotides, wherein each nucleotide comprises a cleavable cyclic loop between two positions of the nucleotide, wherein the cyclic loop comprises reporter moiety corresponding to the identity of the nucleotide. The reporter may be comprised of an amino acid sequence. The cyclic loop may further comprise modifications or arresting constructs for slowing or halting the polynucleotide translocation through a nanopore. Also disclosed are methods of detecting nucleotides and a kit for determining the sequence of nucleotides.

The invention provides a construction method of a bacterial pathogen detection model based on nanopore adaptive sampling and deep learning, and a method, equipment, a medium and a program product for detecting bacterial pathogens by applying the model, and relates to the technical field of nanopore sequencing. The bacterial pathogen detection method comprises the following steps: obtaining a water sample to be detected; and inputting a to-be-detected water sample into the detection model constructed in the invention to obtain a detection result of the to-be-detected water sample. The sensitivity and the accuracy of pathogen detection are obviously improved, and the improvement on prevention and control of water-borne diseases is facilitated.

The invention discloses an electrochemical biosensor of a light-driven nanopore channel and application of the electrochemical biosensor. According to the sensor, a photoresponse nanopore membrane is constructed, an outer surface aptamer probe is combined, asymmetric photothermal gradient driven ion transmission induced by near-infrared light is utilized, and ultra-sensitive detection on a target object is realized by regulating and controlling the charge density of the outer surface. According to the electrochemical biosensor provided by the invention, traditional voltage driving is replaced by asymmetric near-infrared light driving, so that the electrochemical biosensor has a relatively wide detection range and an extremely low detection limit, and high-sensitivity and specific detection on a target object can be realized. The electrochemical biosensor can be used for detecting microcystin-LR (MC-LR), the detection limit reaches 1 * 10 <-7 > mu g / L, the sensitivity is improved by 5 orders of magnitude compared with that of a traditional voltage-driven sensor, and the electrochemical biosensor can be applied to trace pollutant detection in the fields of environmental monitoring, food safety and the like.

The invention discloses a pathogenic microorganism detection system based on nanopore sequencing and intelligent interpretation. The system comprises six modules including a biomolecule electric signal acquisition module and a pathogen characteristic sequence extraction module. Ion current change signals are collected through a nanopore chip array, a pathogen characteristic sequence is extracted through differential coding, a hypergraph topological structure self-adaption building module is used for building a pathogen relation model, a multi-dimensional heterogeneous pathogen parameter fusion analysis module integrates morphological parameters, metabonomics parameters and other multi-dimensional parameters, and the multi-dimensional heterogeneous pathogen parameter fusion analysis module is used for analyzing the multi-dimensional parameters. The optimized hypergraph neural network reasoning module realizes pathogen category prediction, and finally, the nanopore sequencing result intelligent interpretation output module generates a detection report containing pathogen categories, subtypes and risk levels. The system is combined with nanopore sequencing and an intelligent algorithm, pathogenic microorganism detection can be efficiently and accurately completed, the accuracy, reliability and efficiency of detection are greatly improved, and the requirement for rapid and accurate detection of pathogens in practical application is met.

Provided herein are methods and systems for sequencing proteins. One or more methods disclosed herein may use linkers having an amino acid-reactive group and an additional reactive moiety that may be used to couple a polymerizable molecule. The linker may couple to a polymerizable molecule and an amino acid of a peptide and a capture moiety via the polymerizable molecule, followed by cleavage of the amino acid from the peptide. Further processing and analysis may be conducted using, for example, nanopores or nanogaps.

The invention provides a nanoscale metal tip intelligent detector and a use and analysis method thereof. The device integrates a high-precision imaging and positioning module, an intelligent analysis module, an intelligent control interaction module and an automatic execution mechanism. The imaging and positioning module is composed of an optical microscope, a high-resolution CCD camera and a three-axis precision objective table and supports nanoscale morphology feature capture and micron-scale positioning precision. The intelligent analysis module can automatically extract the curvature radius, the taper angle, the surface roughness and other key parameters. The control interaction module is coupled with the automatic execution mechanism, so that automatic control of the detection process and remote storage and optimization of process data can be realized. The device can automatically complete the detection and intelligent analysis of the morphology of the nanoscale metal tip, is compatible with application scenes such as single nanopore / pore channel preparation, scanning probe tip morphology characterization and nano device detection, and remarkably improves the operation efficiency while maintaining high-precision detection.

In one aspect, the disclosed technology relates to nanopore sequencing with a polynucleotide comprising a plurality of nucleotides, wherein each nucleotide comprises a chelating moiety bound to a metal ion. In some embodiments, the chelating moieties are configured for slowing or halting the polynucleotide translocation through a nanopore until a specific stimulus is achieved. In some embodiments, the chelating moieties, as well as any bound metal ions, can serve as reporters for specific nucleotides.