44 results about "NUCLEIC ACID SUBSTANCE" patented technology

The invention provides a tumor cell vaccine as well as a preparation method and application thereof. The preparation method comprises the following steps: acquiring and culturing tumor cells; enabling the cell membrane of the tumor cell to express a targeting antibody by utilizing a gene vector infection or gene editing technology, and enabling the targeting antibody to be used for enabling the tumor vaccine to be combined with the dendritic cell; performing overexpression of tumor specific protein on the cell membrane surface of the tumor cell by using a gene vector infection or gene editing technology; a nucleic acid substance of a target protein is wrapped by lipid nanoparticles to form a vaccine framework, and the nucleic acid substance can enable target cells to express directional chemotactic molecules; wherein the directed chemotactic molecule is used for transferring the dendritic cells to lymph nodes; the target vaccine is obtained by wrapping the vaccine framework with the cell membranes of the tumor cells, lymph node homing of the dendritic cells can be promoted, and the antigen presentation efficiency is increased and the immune effect is enhanced through tumor specific protein carried by the target vaccine and assisted by an immunologic adjuvant.

The invention relates to a quaternary ammonium salt modified strong anion monolithic column, and discloses a monolithic column material, and quaternary ammonium salt groups are grafted on the internal surfaces of pores and channels in the monolithic column material. The quaternary ammonium salt group exists in the form of EP-OCH2CH (OH) CH-N + R1R2R3. The monolithic column prepared by the invention is rich in hydroxyl groups, has excellent compatibility to an elution object containing biomacromolecules, has excellent specific adsorption capacity for nucleic acid substances, and is suitable for enrichment and separation of DNA and / or RNA substances.

The invention discloses a monolithic column material. Primary amine groups or secondary amine groups or tertiary amine groups are grafted on the inner surfaces of pores and channels in the monolithic column material. The primary amine group or the secondary amine group or the tertiary amine group exists in the form of EP-OCH2CH (OH) CH-NR1R2. The monolithic column prepared by the invention is rich in hydroxyl groups, has excellent compatibility to an elution object containing biomacromolecules, has excellent specific adsorption capacity for nucleic acid substances, and is suitable for enrichment and separation of DNA and / or RNA substances.

Provided herein are products and processes for primer extension reactions using chain terminating reagents and enzymes having 3′ to 5′ exonuclease activity, whereby unextended oligonucleotides are digested and extended oligonucleotides are not digested. Also provided herein are products and processes for the detection of the presence, absence or amount of one or more variants of a target nucleic acid species or a plurality of target nucleic acid species comprising primer extension reactions using chain terminating reagents and enzymes having 3′ to 5′ exonuclease activity, whereby unextended oligonucleotides are digested and extended oligonucleotides are not digested.

Disclosed in the present invention is a monolithic column material. Quaternary ammonium groups are grafted onto the inner surfaces of pores and channels of the monolithic column material. The quaternary ammonium group is present in the form of EP-OCH2CH(OH)CH-N+R1R2R3. The monolithic column prepared in the present invention is rich in hydroxyl, has excellent compatibility with an eluted analyte containing biomacromolecules, has excellent specific adsorption capacity for nucleic acid substances, and is suitable for enrichment and separation of DNA and / or RNA substances.

Disclosed is a monolithic column material, primary amine groups or secondary amine groups or tertiary amine groups being grafted on the internal surfaces of pores and channels of the monolithic column material. The primary amine groups, secondary amine groups or tertiary amine groups exist in the form of EP-OCH2CH(OH)CH-NR1R2. A monolithic column prepared by the present invention is rich in hydroxyl groups, has excellent compatibility with elution objects containing biological macromolecules, and also has excellent specific adsorption capacity for nucleic acid substances, making it suitable for enrichment and separation of DNA and / or RNA substances.

The invention discloses a method for preparing chitosan-polyanion nanoparticles based on a microfluidic mixing technology and application. The method comprises the following steps: preparing a water phase 1 as a chitosan acidic aqueous solution, preparing a water phase 2 as a polyanion aqueous solution, and adding a surfactant; by utilizing a microfluidic device and a microfluidic mixer / chip, continuously injecting the two phases according to a set total flow rate and a flow rate ratio, and quickly mixing in a microscale channel, so that the chitosan and the polyanions are subjected to electrostatic compounding and / or ionic cross-linking to form nano-particles; the resulting dispersion is filtered to remove agglomerates and converge the particle size distribution. The aqueous phase 2 may further comprise an anionic drug or active ingredient, preferably a nucleic acid species, to produce supported nanoparticles. The obtained dispersion liquid can be stored at 0-8 DEG C for a short time, or freeze-dried after being added with a cryoprotectant to obtain powder. The process provided by the invention can be continuously amplified, and the particle size consistency and dispersion stability are improved. The nanoparticles are used to deliver the anionic drug or active ingredient into a cell or tissue.

The invention provides a nucleic acid protective agent and a preparation method and application thereof, the nucleic acid protective agent comprises a nucleic acid stabilizing main reagent, an RNA enzyme inhibitor and an antioxidant, the nucleic acid stabilizing main reagent comprises maltodextrin, xylitol and hydroxypropyl-beta-cyclodextrin; the RNA enzyme inhibitor comprises cyclopentanol and tetrahydrofurfuryl alcohol; the antioxidant comprises sodium thiosulfate, sodium pyrophosphate and ascorbic acid. According to the nucleic acid protective agent provided by the invention, free nucleic acid substances including DNA and RNA can be preserved for 2 weeks under the condition of 37 DEG C, preserved for 6 months under the condition of 2-8 DEG C and preserved for 15 months under the condition of-20 DEG C, and the degradation rate is less than 10%.

Disclosed are an oxidation-responsive water-soluble cationic pillararene, and a preparation method and application thereof, the oxidation-responsive water-soluble cationic pillararene is a cyclic small-molecule nucleic acid vector, and is obtained by quaternization reaction of 4-methylborate and tertiary amine-modified pillararene. According to the invention, a delivery vector with a high positive charge density is obtained through small molecule synthesis, the delivery vector is capable of being tightly complexed with negatively charged nucleic acid substances to form a nano-composite, and after the nano-composite enters cells, vector positive charges fall off to release the nucleic acid substances in an oxidation environment, and nucleic acid is efficiently translated and expressed. The delivery vector has the characteristics of simplicity and convenience in synthesis, high delivery efficiency and good biological safety, and has a good application prospect.

The invention belongs to the technical field of biology, and particularly relates to high-throughput multi-mode index marking and sequencing detection on single-cell or subcellular nucleic acid substances. According to the invention, a microbead-coupled particle of a PCR primer containing a bar code is designed, the microbead-coupled particle can be delivered to a plurality of different reaction systems in a single reaction by means of droplet microfluidics and the like, and a specific bar code of the reaction system is added to a target nucleic acid molecule in the reaction system in a high-flux parallel manner; the single cell transcriptome or multi-omics sequencing database building is completed. The method disclosed by the invention has the advantages of large cell flux, simplicity and convenience in operation, low price, low pseudosingle cell rate, low no-load rate of a micro-reaction system, high data quality and strong applicability.

The present invention discloses a lysate for releasing nucleic acid of Mycobacterium tuberculosis and its application, the lysate comprises an enzyme component and a buffer component; wherein the enzyme component comprises lysozyme, colorless peptidase, lytic enzyme, mutanolysin, chitinase, and the buffer component comprises Tween-20, potassium chloride, Tris-HCl, EDTA; the enzyme component and the buffer component are mixed evenly during use. Compared with the commonly used lysate of Mycobacterium tuberculosis, the lysate of the present invention does not require high-temperature heating, mechanical vibration, ultrasound and other additional steps during use, does not require equipment assistance, is easy to operate and has low energy consumption, has low dependence on professional environment and professionals, and does not damage nucleic acid substances, and is more friendly to downstream nucleic acid amplification and detection; the enzyme component in the lysate provided by the present invention has a mild and efficient nucleic acid release effect for the cell wall of Gram-positive bacteria, which is not only applicable to Mycobacterium tuberculosis, but also has universality for Gram-positive bacteria.

The invention provides an extraction-free rapid nucleic acid releasing agent and application thereof, and relates to the technical field of molecular biology, and the extraction-free rapid nucleic acid releasing agent comprises 10-100 mM of strong alkali, a surfactant with the mass-volume ratio of 0.1%-2%, 10-100 mM of a buffer solution, 0.1-2 mM of a metal ion chelating agent, 10-100 mM of a lysis enhancer and 50-100 mM of a PCR enhancer I. Cells or virus particles can be fully split, nucleic acid substances are released, and independent inactivation is not needed; the method has the advantages of strong cracking capability, short cracking time and simple steps, and can adapt to a rapid detection program.

The invention relates to the technical field of exosome carrying, in particular to an exosome functional compound and application thereof.The exosome functional compound at least comprises exosomes and nucleic acid substances, the exosomes are derived from animals, plants and microorganisms, the nucleic acid substances are selected from DNA, RNA and fragments thereof, each (1-3) * 10 < 12 > exosomes contain 1-3 mg of nucleic acid substances, and each (1-3) * 10 < 12 > exosomes contain 1-3 mg of nucleic acid substances. Nucleic acid substances are wrapped in exosomes to obtain an exosome functional compound, then the exosome functional compound is mixed with a soluble matrix material and auxiliary materials, the soluble microneedle of the exosome functional compound is formed through an air exhaust process, effective substances can be comprehensively delivered to a corium layer, and the beautifying and anti-aging effects are achieved.

1. Name of the product in this design: Reagent Reaction Tube. 2. Purpose of this design: To hold and seal the reaction system consisting of the nucleic acid substance to be tested and the PCR amplification reagent, and to allow the reaction to take place within the system. 3. The key design feature of this product is its shape. 4. The image or photograph that best illustrates the design's key points: a 3D view of the assembled state.

Disclosed in the present invention is a monolithic column material, which has pyridine groups grafted onto the internal surfaces of pores and channels thereof. The pyridine groups are present in the form of EP-OCH2CH(OH)CH-SPy. The monolithic column prepared in the present invention is rich in hydroxyl, has excellent compatibility with elution objects containing biomacromolecules, and also has excellent specific adsorption capacity for nucleic acid substances, thereby being suitable for the enrichment and separation of DNA and / or RNA substances.

To provide methods for improving cost efficiency, recovery efficiency, and other performance metrics as well as overall process speed for Duplex Sequencing and other sequencing applications for achieving high accuracy sequencing reads.SOLUTION: The present technology relates generally to methods and compositions for targeted nucleic acid sequence enrichment, as well as uses of such enrichment for error-corrected nucleic acid sequencing applications and other nucleic acid sequence interrogations. In some embodiments, provided methods provide non-amplification based targeted enrichment strategies compatible with use of molecular barcodes for error correction. Other embodiments provide methods for non-amplification based targeted enrichment strategies compatible with direct digital sequencing (DDS) and other sequencing strategies (e.g., single molecule sequencing modalities and interrogations) that do not use molecular barcoding.SELECTED DRAWING: Figure 1

The invention relates to a method for preparing a single-step staining reagent for mammal blood detection and detecting visible components. The single-step staining reagent comprises an acid-base regulator, osmotic pressure regulating salt, an accelerating agent, a dye, a protective agent and water, the osmotic pressure regulating salt is used for regulating the osmotic pressure environment in the cell staining process, and regulating the osmotic pressure of the microscopic examination sample to 150-230 mosmol / kg; the pH value of the cell staining process is regulated by the pH regulator, and the pH value of the microscopic examination sample is regulated to 7.0-8.0; the accelerating agent is used for promoting the affinity of the dye and nucleic acid substances in cells; the dye is neomethylene blue, a dyeing substance is provided in the dyeing process, and the content of the neomethylene blue in a microscopic examination sample is 4.0 * 10 (-3) g / L to 5.0 * 10 (-1) g / L; the protective agent can enable protein in a cell membrane to be chemically crosslinked in the dyeing process, a three-dimensional network skeleton is formed to protect the cell membrane from being broken, and a dyeing substance can pass through the three-dimensional network skeleton.

The invention discloses a Zika virus detection target as well as a rapid detection method and a kit thereof, and particularly relates to a nucleic acid substance obtained from an exosome released from cells after human umbilical vein endothelial cells are infected by Zika virus, which can be used as a target for early infection detection. The invention also discloses a rapid detection method established aiming at the target and a kit for rapidly detecting Zika virus infection. According to the method, a probe and a nucleic acid simulant are designed according to a miR-216b-5p sequence with the most significant relative quantitative increase after human umbilical vein endothelial cells are infected with Zika viruses; a fluorescent group is designed at the 5'end of the probe, after adsorption quenching is conducted through carboxylated graphene oxide molecules, the fluorescent group is combined with target nucleotide and then separated from a quenching material, a fluorescent signal is sent out again, early infection of the Zika virus can be early warned by detecting the signal, and the probe has the advantages of being high in sensitivity stability and high in specificity and can be used for detecting early infection of the Zika virus. The method has great practical application value.

Provided a technology platform for the delivery of nucleic acids, centered around a new chemical entity known as Fatty Acid Derivatives (FADS). FADS can covalently or non-covalently bind to nucleic acid substances, thereby enhancing their stability and effectiveness.

The invention provides a fatty acid derivative for delivering nucleic acid substances and a synthesis method thereof. The fatty acid derivative is used for modifying nucleic acid substances such as long-chain nucleic acid, oligonucleotide or double-chain small nucleic acid. The fatty acid derivative can be covalently or non-covalently bound with the nucleic acid substance, so that the in-vivo efficacy of the nucleic acid substance is enhanced, the side effect is reduced, and the storage stability is improved.

The invention provides a host nucleic acid removal method for high-throughput sequencing, and belongs to the field of pathogenic microorganism detection. By limiting the types and use concentrations of reagents, host nucleic acid substances are ensured to be fully removed. In a high host background sample, the method is suitable for tissues and blood, background host nucleic acid can be effectively reduced, and the detection rate and the detection quantity of pathogenic microorganism gene sequences are improved.

This invention belongs to the field of fluorescent dyes, specifically relating to a method for preparing a biostable compound and its applications. It exhibits high stability in both the extracellular and intracellular environments at physiological pH and temperature, making it suitable for biochemical detection and live-cell imaging. This invention also discloses that the probe can detect intracellular and extracellular nucleic acids with high specificity and responsiveness, and its application in the highly sensitive detection of mitochondrial RNA-DNA hybrids in live cells. The fluorescent probe of this invention features strong color development, good photostability, and good biostability. Furthermore, the preparation process of the fluorescent probe is simple and inexpensive, and its stable structure facilitates storage, offering broad application potential in lysosomal and mitochondrial biological function research.

This invention discloses an oxidation-responsive cationic water-soluble pillararene, a method for producing the same, and its use. The oxidation-responsive water-soluble cationic [A2]pilarene is a cyclic low-molecular-weight nucleic acid vector obtained by a quaternization reaction of a 4-methylboronic acid ester with a tertiary amine-modified pillararene. This invention achieves the synthesis of a low-molecular-weight delivery vector having a high positive charge density, which can tightly bind to negatively charged nucleic acid substances to form nanocomplexes. Upon entering a cell, the positive charge of the vector is released in an oxidative environment, the nucleic acid substance is released, and nucleic acid information is efficiently expressed. This delivery vector is easy to synthesize, has high delivery efficiency, and is biologically safe, and is expected to have promising future applications.

The present application relates to the technical fields of biological medicine and gene therapy, and particularly relates to a protonatable cationic lipid, a nucleic acid delivery system and application thereof. The protonatable cationic lipid has a structure as shown in formula (I), and contains a protonatable group such as a hydroxyl group or an amino group at the end. The protonatable cationic lipid can form a lipid nanoparticle for loading various nucleic acid substances. The lipid nanoparticle nucleic acid delivery system has high cell transfection efficiency in vitro, and can be targeted to organs such as liver in vivo to achieve efficient gene editing. The lipid nanoparticle constructed by the protonatable cationic lipid provided by the present application has high encapsulation efficiency, good stability and excellent delivery efficiency, and has a wide application prospect in the fields of gene therapy and cell engineering.

The invention provides a bioactive nanometer device and a preparation method and application thereof, the bioactive nanometer device comprises mBSG-NH2 and a nucleic acid substance loaded on the mBSG-NH2, and the mBSG-NH2 is nanometer mesoporous borosilicate bioactive glass with the surface modified with-NH2. According to the invention, the degradability of the nucleic acid delivery carrier and the protection effect on loading, enzymolysis / thermal inactivation and the like of nucleic acid substances can be improved, and meanwhile, the transmembrane delivery and intracellular lysosome escape performance of the nucleic acid substances can be improved.

The invention discloses collagen nano-microspheres as well as a preparation method and application thereof. The preparation method comprises the following raw materials in percentage by weight: 0.01 to 2 weight percent of collagen, 0.1 to 8 weight percent of nucleic acid substances, 0.001 to 5 weight percent of electropositive substances, 0.1 to 7 weight percent of auxiliaries and 78 to 99.78 weight percent of water, wherein the total mass of the raw materials is 100 weight percent; the method comprises the following steps: mixing a nucleic acid substance with partial mass of water, performing high-pressure homogenization or microjet treatment, performing low-temperature curing to obtain nucleic acid semi-solid gel, and heating and melting to obtain a phase A; dissolving collagen in the phase A to obtain a phase B; mixing the phase B and the phase C, performing high-pressure homogenization or micro-jet treatment, and adding an auxiliary agent to form collagen nano-microspheres; wherein the electropositive substance is dissolved in the balance of water in the raw materials to obtain a phase C; the collagen microspheres can improve transmembrane transport of recombinant collagen and bioavailability, and meet application requirements of the collagen microspheres in products such as transdermal drug delivery drugs.

1. The name of this design product: Nucleic acid extractor. 2. The use of this design product: It is used for extracting nucleic acid substances from biological samples and is applicable to clinical gene testing laboratories, disease prevention and control centers, research institute laboratories, and medical college laboratories. 3. The design key points of this design product: lie in the combination of shape and pattern. 4. The picture or photo that best shows the design key points: Perspective view 1.

This application relates to multiplexed methods for the identification and quantification of minor alleles and polymorphisms. Products and methods are provided for detecting the presence of minor nucleic acid substances in a sample containing a mixture of minor nucleic acid substances and one or more major nucleic acid substances, wherein the amount (frequency or copy number) of the minor nucleic acid substances is less than the amount of the major nucleic acid substances. Some methods involve amplifying the mixture and extending the resulting amplicons with a chain terminator and an extension primer that hybridizes specifically to the amplicons, wherein the chain terminator specific to the major nucleic acid substances has a lower concentration than the chain terminator specific to the minor nucleic acid substances. A higher concentration of the chain terminator specific to the minor nucleic acid substances, relative to the chain terminator specific to the major nucleic acid substances, improves the detection limit (sensitivity) for detecting minor nucleic acid substances present in the mixture at low frequency or copy number. Additionally, the signal generated from the extension product of the major nucleic acid substance amplicons can be used as a positive control and allows for the quantification of the minor nucleic acid substances relative to the major nucleic acid substances in the mixture.

1. The name of the design product: nucleic acid detection analyzer. 2. The use of the design product: the design product is used for extracting, amplifying and detecting analysis of nucleic acid substances in biological samples. 3. The design points of the design product: in shape. 4. The picture or photo that best indicates the design points: perspective view 1.

The invention discloses a protective agent for nucleic acid substances and application of the protective agent. The protective agent comprises nanoparticles, trehalose, mannitol and bovine serum albumin, wherein the nanoparticles are provided with cavities capable of wrapping nucleic acid substances. The protective agent can maintain the structure of nucleic acid substances in the high-temperature drying process and reduce the water content at the same time, and normal-temperature storage and transportation of the nucleic acid substances are achieved; the stability of nucleic acid substances at lower concentration can be improved, and the sensitivity or the detection rate of the PCR detection kit is improved.