340 results about "Nanopore" patented technology

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

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 sensea 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.

Provided herein are methods of interrogating spatial gene expression in a sample using substrates having sequencing pores. The disclosed methods allow for spatial analysis of analytes from biological samples using a pore-based sequencing approach and without the need for a barcoded spatial array. In some embodiments, an analyte or intermediate agent thereof, is released from a biological sample and directly sequenced by traversing through pores of a sequencing array including a plurality of pores, e.g., nanopores. In some embodiments, light or other stimuli is used to release an analyte or intermediate agent thereof from a specific region of interest in the biological sample followed by sequencing using sequencing array including a plurality of pores.

Example systems and methods for de novo sequencing of DNA or DNA-like sequences using Viterbi-like correlation analysis are described. A sequencing system receives the read data for multiple copies of a DNA strand from a sequence reader, such as a nanopore reader. The sequencing system generates a convolutional matrix based on one copy and a reference matrix based on another copy and uses them to generate a correlation matrix. A most likely path through the correlation matrix is determined to identify and correct errors between the two copies.

The invention provides a glass nanopore probe based on an aptamer and application of the glass nanopore probe in dopamine detection, and belongs to the technical field of electrochemical sensing and scanning probe microscopy. The sensor comprises: a glass nanopore probe, the inner wall of which is modified with a gold layer; a cDNA (complementary deoxyribonucleic acid) single chain is fixed on the gold layer through a gold-sulfur bond; a part of the sequence of the dopamine aptamer is hybridized and combined with the cDNA single chain; wherein the combination of the dopamine and the dopamine aptamer causes the change of an ion current rectification signal of the glass nanopore probe, so that the detection of the dopamine is realized based on the change of the ion current rectification signal of the glass nanopore probe. The invention provides a valuable scheme for developing an aptamer nanopore scanning electrochemical sensor for single cell analysis and researching neurotransmitter-related diseases.

The invention discloses a species identification kit for animal mitochondrial bar code multiplex PCR amplification and nanopore sequencing, and relates to the technical field of species identification. The species identification kit comprises optimized primer pairs aiming at four hyper-variable genes (Cytb, COI, 16S rRNA and 12S rRNA) of mitochondria, by introducing R, Y, D and other degenerate bases at key sites, the binding capacity of the primers and template DNA of different species is remarkably improved, the problems that an existing universal primer is prone to mismatch, low in efficiency and even failed in amplification during cross-species amplification are solved, and the species identification kit has the advantages of being high in specificity, high in specificity and high in specificity. The method can be used for species identification of a mixed DNA sample containing at least two different animal species sources. The species identification primer combination is applied to preparation of an animal species identification kit, and the detection coverage range of the kit is expanded. According to the animal species identification method provided by the invention, a multiple PCR amplification system and a nanopore length reading sequencing technology are combined, congeneric sibling species with highly similar sequences can be distinguished, and the method has good accuracy.

A combination of lipid-binding molecules and / or lipid-binding proteins with lipid components (i.e., mispids) is provided for use in modifying the interaction between target molecules and lipid membranes. This includes, for example, the use of lipid-binding molecules and / or mispids to improve the sequencing efficiency and throughput of nanopore-based sequencing systems. To sequence target molecules such as nucleic acid sequences or nucleic acid substitute polymers derived therefrom, lipid-binding molecules and / or their mispids are combined with the target molecules. The mixture is then applied to a nanopore-based sequencing chip. The target molecules are then sequenced in the presence of lipid-binding molecules and / or nanodiscs, thereby improving the capture, arrival time, and effective concentration of the target molecules across the chip membrane. Such improved efficiency is particularly beneficial, for example, when the concentration of the target molecule is low.

The utility model discloses an integrated solid-state nanopore biological detector, which relates to the technical field of biological detection and comprises a fluid pool shell, a solid-state nanopore silicon nitride chip and a PCB (printed circuit board), a sample injection hole is arranged on the fluid pool shell, the solid-state nanopore silicon nitride chip is arranged in the fluid pool shell, and the solid-state nanopore silicon nitride chip is arranged in the PCB. A silicon nitride window corresponding to the solid-state nanopore silicon nitride chip is formed in the fluid pool shell, a PCB (Printed Circuit Board) is fixed in the fluid pool shell, an RFID (Radio Frequency Identification Device) storage chip, a magnetic connector and silver electrodes are arranged on the PCB, and the silver electrodes are positioned on two sides of the silicon nitride window. According to the utility model, the RFID storage chip is arranged, so that the key information of the biological detector is stored and traced, and the problem of information management disorder in the prior art is solved; the RFID storage chip can store key information of the biological detector for a long time, and the integrity and traceability of the data are ensured.

To provide a system including novel nanopores having different structures and recognition sites that improve sequencing accuracy and / or provide different error profiles.SOLUTION: Provided are systems relating to the field of nanopores and their use in various applications, e.g., analysis of biopolymers and macromolecules, typically by making electrical measurements during translocation through the nanopore, comprising a funnel-shaped proteinaceous nanopore comprising alpha-helical pore-forming toxins that are members from the actinoporin family of proteins, more particularly Fraga Ceatoxin C (FraC), a variant FraC, a FraC paralog or a FraC homolog.SELECTED DRAWING: None

The invention relates to means and methods for analysis of target analytes using nanopore-based sensors, more in particular to methods, nanopore systems and devices for single-molecule profiling of polymers, e.g. polypeptide or polysaccharides. Provided is a method for translocating a non-nucleic acid based polymer analyte through a nanopore, the nanopore being comprised in a membrane separating a fluidic chamber of a nanopore system into a cis side and a trans side, comprising adding the analyte to the cis side of and allowing for translocation, wherein the nanopore system has a cis to trans electro-osmotic force (EOF) resulting from a net ionic current flow cis to trans, preferably wherein the cis to trans EOF results from a net ionic current flow cis to trans over total ionic current flow of greater than 0.2 or less than −0.2.

The invention discloses a detection system and method for identifying mycobacteria and drug resistance genes of the mycobacteria based on nanopore sequencing, and relates to the field of biological medicine, the detection system comprises a specific targeted enrichment module, a nanopore sequencing module and a biological information analysis module; the specific targeted enrichment module comprises a probe combination, and the probe combination covers a mycobacterium tuberculosis complex conservative identification gene, species-specific genes of common nontuberculous mycobacteria and mycobacterium leprosy, and full-length or partial sequences of drug resistance related genes in a targeted manner; by utilizing the characteristics of nanopore length reading length and real-time sequencing and a tuberculosis specific targeted enrichment strategy, accurate identification of a mycobacterium tuberculosis complex group, synchronous typing of 42 mycobacteria, analysis of 24 drug-resistant genes, and efficient detection of structural variation and low-abundance heterogeneity drug-resistant mutation are realized, the detection period is shortened, the detection cost is reduced, and the detection efficiency is improved. And a comprehensive and reliable technical basis is provided for accurate diagnosis and treatment of mycobacterium infection.

Provided are methods, compositions, and systems for the detection of a target analyte. Also provided are methods, compositions, and systems for determining the concentration one or more target analytes in fluid solution. The compositions include nanopore conjugates in which a nanopore protein monomer is joined to a capture tag. Tethered to the nanopore protein conjugate is an analyte ligand directed to a specific analyte. When a voltage is applied across a nanopore assembly including the nanopore conjugate, the nanopore captures the capture tag at a given capture rate. In the presence of the analyte to the analyte ligand, however, the capture rate of the capture tag changes, thus permitting detection of the analyte by the nanopore assembly. Further, based on the capture rate associated with binding between the analyte and the analyte ligand, the concentration of the analyte can be determined using association / dissociation kinetics.

The invention relates to the technical field of electrochemical luminescence sensing, and discloses an electrochemical luminescence immunosensor for detecting interleukin-1 beta, a preparation method and application thereof, the electrochemical luminescence immunosensor comprises a working electrode, a reference electrode and a counter electrode; the working electrode comprises a substrate electrode and an ordered mesoporous silica film (SNF) modified on the surface of the substrate electrode; a composite nano-enzyme containing cobaltosic oxide nanoparticles and nitrogen-doped graphene quantum dots is loaded in a nano-pore channel of the SNF, the outer surface of the SNF is modified with an epoxy group, and an anti-interleukin-1beta antibody is covalently immobilized through the epoxy group; a detection system of the electrochemical luminescence immunosensor is a luminol / dissolved oxygen system. According to the electrochemical luminescence immunosensor, a luminol / dissolved oxygen system signal can be remarkably enhanced, and high-sensitivity and accurate quantitative detection of interleukin-1 beta is realized under a neutral condition.

The invention relates to the technical field of biomedical detection, and discloses a system for detecting extracellular matrix free metalloproteinase and a soluble receptor. Comprising a polypeptide substrate nanopore sensing module used for detecting the enzyme activity of free metalloproteinase in an extracellular matrix; the electrophoretic analysis module is used for detecting a soluble receptor in an extracellular matrix, applying an electric field to a tumor microenvironment sample, determining the inter-terminal resistance of the sample by using an inter-terminal resistance detection device, and detecting the soluble receptor by comparing the resistance change according to the characteristic that the existence of the soluble receptor causes the additional resistance increase of the sample; and the data processing and judging module is electrically connected with the polypeptide substrate nanopore sensing module and the electrophoretic analysis module respectively, and is used for receiving the free metalloproteinase activity detection result and the soluble receptor detection result, and judging the time interval of cellular immune escape according to a preset rule in combination with cellular morphology observation information.

The invention provides a nanopore probe and application thereof, the nanopore probe comprises a polypeptide, the carboxyl terminal of the polypeptide is connected with a steric hindrance tag, the amino terminal of the polypeptide is connected with polyaspartic acid, and the polypeptide has a restriction enzyme cutting site specifically recognized by target protease. The method can prolong the retention time of the polypeptide in the nanopore, enhance the signal discernibility and improve the signal stability, thereby realizing stable and high-precision quantitative detection of the protease activity.

Provided is an electrode catalyst having excellent catalyst activity, which contributes to the low cost of a PEFC. The electrode catalyst includes a hollow carbon carrier having nanopores with a pore diameter of 1 to 20 nm, and a plurality of catalyst particles supported on the carrier. The catalyst particles contain Pt (0 valence), are supported inside and outside the nanopores of the carrier, and in a case where analysis of the particle size distribution of the catalyst particles is performed using a three-dimensional reconstruction image obtained by electron tomography using STEM, the proportion of the catalyst particles supported inside the nanopores is 50% or more, and at least one nanopore having the shape of a continuous communication hole is formed in a cubic image having one side of 20 to 50 nm obtained from the three-dimensional reconstruction image of the catalyst block.

Provided herein are methods of characterising a target polypeptide as it moves with respect to a nanopore. Also provided are related kits, systems and apparatuses for carrying out such methods.

The application discloses a two-dimensional metal organic framework nanopore sensor and a preparation method and application thereof. The two-dimensional metal organic framework nanopore sensor comprises a two-dimensional metal organic framework film with first nanopores, a silicon nitride film with second nanopores and a silicon substrate which are sequentially stacked, the two-dimensional metal organic framework film is formed through liquid-liquid interface force self-assembly and is transferred in situ to the silicon nitride film to form. The two-dimensional nanopore film material adopted by the application is a metal organic framework of a porphyrin organic ligand, the hydrophobic porphyrin ligand has high affinity with DNA molecules and can interact with the DNA molecules, so that the speed of the DNA passing through the two-dimensional nanopore is slowed down, the problem of fast pore passing speed when the two-dimensional nanopore sensor detects and distinguishes short-chain DNA is solved, and the time resolution capability of the two-dimensional nanopore is greatly improved under the premise of guaranteeing high spatial resolution of the two-dimensional nanopore.

The invention provides a nanopore dispersion strengthened copper material and a preparation method thereof. The material is obtained through a two-step method of firstly oxidizing and then reducing: firstly, adding an oxygen element into copper, namely preparing a precursor with a Cu-Cu2O two-phase structure from copper and copper oxide, and adopting a powder coating-laser powder bed melting or electric arc melting-solidification process; and then performing thermal reduction in a reducing atmosphere, so that cuprous oxide in the precursor is reduced and converted into closed nanopores in situ. The obtained material is composed of a copper matrix with micron-sized crystal grains and nano-sized holes which are uniformly and dispersedly distributed. According to the method, the atomic ratio of Cu to O, the solidification cooling speed and the reduction temperature / time are adjusted, and the volume ratio of the Cu2O phase in the precursor is adjusted through high-temperature annealing aftertreatment, so that the porosity and the pore diameter are adjusted and controlled. According to the method, heterogeneous elements are not introduced, the process is universal, the cost is friendly, and the obtained material has high strength, high conductivity and excellent high-temperature structural stability.

The application discloses a preparation method of Al / NH4CuF3 metastable intermolecular complex. The method is characterized in that: firstly, NH4CuF3 with a nano-network structure is prepared through a solvothermal reaction; then, n-Al is filled into the nanopore channels of the NH4CuF3 in a manner of ultrasonic-assisted dispersion, so as to form n-Al / NH4CuF3 MICs with good dispersity. The n-Al / NH4CuF3 MICs prepared by the method can etch a passivation layer and reduce the reaction energy barrier of the aluminum powder and an oxidizing agent, depending on the low-temperature decomposition of the NH4CuF3 to release F and the thermal decomposition of the NH4CuF3 to generate HF. In addition, a large amount of gaseous products are generated by the decomposition of the NH4CuF3, so that the generation of a reaction sintering phenomenon is relieved, and the combustion efficiency of the n-Al is improved.

The present disclosure relates to the field of protein detection technology, and discloses a protein detection method based on a combined nanopore structure, comprising: a surface of a sandwich film chip is processed to form a nano-through hole, and a “hole-cavity-hole” structure is prepared. The “hole-cavity-hole” structure is driven by the electric field of the functional protein to detect the protein. The present disclosure adopts the above-mentioned protein detection method based on a combined nanopore structure, which can effectively overcome the problem of protein detection reading length caused by the limitation of nanopore space. Meanwhile, in contrast to methods that embed functional protein complexes within a phospholipid bilayer, integrating functional proteins directly into solid nanopores combines the advantages of both biological and solid systems. This approach not only relaxes the stringent environmental demands placed on the sensor but also enhances its resolution and stability.

The invention discloses a carbohydrate molecule sequencing method based on glycosidase and nanopores and a system and application thereof. The invention also discloses a nanopore mutant protein and application thereof. The invention also discloses a glycosequencer containing the nanopore protein, a sequencing method, a system and application thereof. The invention also discloses a data processing method and a processing system for glycosidase hydrolysis assisted nanopore sugar sequencing, a computer related product and the like. According to the method, the detection limit can be remarkably reduced, the sensitivity can be improved, the accuracy of sugar sequencing can be improved, the analysis accuracy exceeds 98%, the sensitivity is improved by more than 50 times, the time is saved by more than 5 times, the analysis period is short, the sequencing process is simplified, and the cost is reduced. And a new technical approach is provided for rapid analysis of complex sugar sequence information.

Methods and devices for nanopore-based biomarker detection that utilize immunocapture, isothermal rolling circle amplification, and sequence-specific fragmentation of the product to release multiple DNA reporter molecules for nanopore detection. These DNA fragment reporters produce sets of nanopore signals that form distinctive fingerprints. This fingerprint signature therefore allows the identification and quantification of biomarker analytes.

Provided herein are Mycobacterium smegmatis porin nanopores, systems that comprise these nanopores, and methods of using and making these nanopores. Such nanopores may be wild-type MspA porins, mutant MspA porins, wild-type MspA paralog porins, wild-type MspA homolog porins, mutant MspA paralog porins, mutant MspA homolog porins, or single-chain Msp porins. Also provided are bacterial strains capable of inducible Msp porin expression.

The invention provides a three-stage translocation model for revealing Aerolysin nanopore fingerprint characteristics, a mathematical framework related to the nanopore fingerprint characteristics (charge-to-mass ratio, resistance-to-mass ratio and other parameters) is established by decomposing a charged single-molecule translocation total process into three stages of pore entry, perforation and pore exit, and a single-molecule-level fingerprint characteristic extraction technology is established. And explaining the translocation attribute of the single molecule by using fingerprint characteristics. Theoretical and experimental analysis proves that the molecular charge-to-mass ratio is positively correlated with the metathesis speed, the molecular metathesis speed is dominated by the charge-to-mass ratio during hole entering, and the acceleration phenomenon of molecules in the hole exiting stage is predicted. The invention provides a framework for quantitative characterization, modeling and prediction of nanopore fingerprints in biosensing applications.

This disclosure pertains to the field of gene sequencing technology and discloses the KphPif1 helicase and its applications. This disclosure relates to a modified helicase, a polynucleotide encoding the helicase, a recombinant expression vector, and a construct capable of expressing the modified helicase and used to construct recombinant cells expressing the helicase. The modified helicase disclosed herein allows target polynucleotides to pass through nanopores in a controllable and stepwise manner, stabilizing polynucleotide movement and preventing detachment from the polynucleotide. Furthermore, it features a wide current range, high decoding accuracy, fast perforation speed, and high sequencing efficiency.

The invention relates to the technical field of biology, and discloses a linker element and a method for constructing an oxford nanopore direct RNA sequencing library by the linker element, and the linker element comprises a partial double-strand dump structure formed by annealing a first nucleotide strand S and a second nucleotide strand AS; the first nucleotide chain S sequentially comprises a ligase recognition motif region, a sample tag sequence region, a first universal sequence region and a second universal sequence region in the 5 '-3' direction; the second nucleotide AS sequentially comprises a third universal sequence region, a first universal sequence reverse complementary region, a sample tag sequence reverse complementary region, a ligase recognition motif reverse complementary region and a polyadenylate tail recognition region according to the 5 '-3' direction. According to the method, RNA can be efficiently utilized, the high sequencing data utilization rate and high resolution uniformity are achieved, the technical problems that an existing Oxford nanopore direct RNA sequencing technology cannot effectively utilize initial RNA, and a plurality of library samples cannot be effectively split on the same chip are solved, and the economic and time cost of a practical application end is reduced.

The invention discloses a reversible dynamic aperture solid nanopore array based on a temperature-sensitive phase change layer and a preparation method and application of the reversible dynamic aperture solid nanopore array, and belongs to the technical field of single molecule detection and nanostructure regulation, the array introduces the temperature-sensitive phase change layer into the inner wall of a solid nanopore, and when the local temperature crosses the phase transition point of the temperature-sensitive phase change layer, reversible volume change is generated; therefore, the effective aperture of the nanopore is continuously adjustable in a sub-nanometer scale range. An independent micro heating unit and an independent temperature sensing unit are integrated below each nanopore in the array, and single-pore-level accurate temperature control adjustment is achieved in a row-column addressing mode. In combination with ionic current monitoring and an external control module, the aperture can be set in real time according to the size requirements of different target molecules, and dynamic aperture regulation and control with a programmable structure, quick response and good repeatability are realized. The array structure provided by the invention is suitable for single molecule level detection of objects such as multiple types of small molecule pollutants, tumor-related biomarkers, biomacromolecules and the like, and has the advantages of high sensitivity, high flux and wide applicability.

The invention relates to the technical field of isotope batteries and discloses a nanopore-containing power generation assembly. The nanopore-containing power generation assembly comprises a transduction layer, the loading layer is arranged on the surface of the transduction layer in advance, and the loading layer is provided with preset nanopores. The technical problem that in the preparation process of the radioactive source in the isotope battery, due to the fact that the hydrogen absorption temperature of a hydrogen storage material is high, large thermal mismatching exists between a transducer device and the hydrogen storage material, growth stress is generated in the phase change process, and consequently the transducer device may have serious structural damage can be solved; the loading layer arranged on the surface of the transduction layer in advance is adopted, and the loading layer is provided with the nanopore structure, so that when the loading layer serves as a basic structure for preparing the isotope power generation assembly, for example, nanopores are filled with radioactive gas such as tritium, the loading layer structure is stable, and the isotope power generation assembly is prepared. The probability that the energy conversion layer structure is damaged due to growth stress generated in the phase change process can be reduced, and then it is guaranteed that the structure of the prepared isotope power generation assembly is stable.