98 results about "NUCLEIC ACID SUBSTANCE" patented technology

The invention provides a method and a system for digital quantitative analysis of nucleic acid amplification based on micro-droplet. The method comprises the following steps: preparing a to-be-detected nucleic acid amplification reaction liquid which includes a to-be-detected nucleic acid template, a reaction buffer water solution, deoxyribonucleoside triphosphate, a primer, polymerase and a product marking substance; loading the prepared to-be-detected nucleic acid amplification reaction liquid in a micro-pipeline of which the two ends are both provided with an opening, wherein the micro-pipeline is arranged above an open container, and the open container contains an oily liquid containing a surfactant; enabling the opening of one end of the micro-pipeline to do up-and-down reciprocating vibration on the surface of a liquid in the open container or do leftward-and-rightward reciprocating vibration below the liquid surface so as to generate a plurality of droplets which are flatly laid at the bottom of the open container; performing nucleic acid amplification reaction on the plurality of droplets in the open container; acquiring product signals generated after the nucleic acid amplification reaction is ended, and performing quantitative analysis on the nucleic acid template. The digital nucleic acid amplification analysis method can be used for implementing quantitative detection on low-concentration nucleic acid substances in a micro-system and is simple and convenient to operate, high in efficiency and low in cost.

The invention discloses a method and application for preparing a tissue engineering corneal carrier stent by utilizing fresh porcine cornea. The method comprises the following steps: cutting a fresh porcine cornea for swelling, repeatedly freeze thawing and breaking cells, removing residual nucleic acid substances through DNA-RNA enzyme treatment, cross-linking, airing, and finally performing irradiation sterilization through ionization rays, thereby obtaining the product. The method is applied to carrier stents of tissue engineering corneal epithelium, stroma, endothelium, front board layer half cornea, rear board layer half cornea and full-layer board layer half cornea, so that the requirement on batch production of tissue engineering corneas is met. The carrier stent serves as a substitute of human corneal stroma to be used for clinical transplant and treatment of un-accumulated whole layer keratohelcosis. The method is applied to large-scale production and clinical popularization and application. The prepared carrier stent has the characteristics of high transparency, dense structure, high mechanical property, high biocompatibility with human corneal seed cells and the like. Therefore, the method is suitable for batch production, so that the requirements on lots of carrier stents for tissue engineering corneas are met.

The invention relates to a lumbricus extract possessing antithrombotic effect, which takes lumbricus decoction pieces as a raw material. The preparation method comprises the following steps: coarse cracking the raw material of the lumbricus decoction pieces, immersing for 4-5 hours by adding 5-15 times water by weight, adding immersion liquid together in a pulverizer to crush by a wet method so as to obtain slurry with average particle fineness of D50<=100 mum, centrifuging and taking a supernatant, passing through a ceramic membrane with average aperture of 0.45 mum, refrigerating a filtrateat the temperature less than or equal to minus 20 DEG C for 2 hours, and then refrigerating and drying the filtrate under the condition of minus 4 DEG C-minus 6 DEG C of temperature and 0.1mbr-0.05mbr of absolute pressure. The lumbricus extract is capable of extracting amino acid and nucleic acid substances, and keeping the activity of effective chemical ingredients of protein and polypeptide with maximum limit. The lumbricus extract has dual effects of anticoagulation and thromboclasis, animal experiments show that the lumbricus extract has the advantages of good antithrombotic effect and simple extraction method with high efficiency.

The invention discloses a virus detection system and a micro-fluidic chip thereof. The micro-fluidic chip comprises a liquid storage unit used for storing a liquid used or generated in the detection process, a nucleic acid purification unit communicated with the liquid storage unit and used for purifying a sample liquid to obtain a nucleic acid substance, and an amplification detection unit comprising a main pipeline, a plurality of reaction tanks and a plurality of branch pipelines, wherein the amplification detection unit is communicated with the nucleic acid purification unit through the main pipeline, the reaction tanks are communicated with the main pipeline through the branch pipelines, and the reaction tanks are used for amplifying and detecting nucleic acid substances of differentsample solutions. The multifunctional micro-fluidic chip integrates nucleic acid extraction, nucleic acid purification and nucleic acid amplification and detection. The requirements of all steps of nucleic acid amplification and detection can be automatically met on the premise of one-time sample introduction, and a plurality of reaction tanks are arranged so that amplification and detection of aplurality of sample solutions can be achieved at the same time and the detection efficiency is high.

The invention discloses a nucleic acid substance carrier containing a degradable imine linkage. The structural formula of the nucleic acid substance carrier containing the degradable imine linkage is described in the specification, wherein x is 1-20, y is 1-20, and n is 1-20. The invention also discloses a preparation method of the nucleic acid substance carrier containing the degradable imine linkage, an application in nucleic acid medicine delivery and a complex. Compared with the prior art, the nucleic acid substance carrier which contains an imine linkage structure and can degrade low-molecular-weight PEI (polyether imide) derivatives is simple in structure and easy to synthesize, shows higher transfection activity in different cell strains and has lower cell toxicity, so that the nucleic acid substance carrier is a high-efficiency and low-toxicity gene delivery carrier.

The invention provides a micro-volume cell nucleic acid amplification method which comprises the following steps: a) putting micro liquid droplets with a small amount of cells inside into a small-sizecontainer with an oil phase supplied in advance, and performing centrifugation to settle down the micro liquid droplets; b) putting cell lysis buffer droplets into the small-size container through micro-volume injection below the liquid level of the oil phase, performing centrifugation to settle down the cell lysis buffer droplets, and fusing the cell lysis buffer droplets with cell droplets so as to achieve splitting of cells and release of nucleic acid substances; c) adding splitting termination droplets through micro-volume injection, performing centrifugation, fusing the splitting termination droplets with the split cell droplets, and neutralizing or terminating the splitting reaction; d) adding one or more amplification reaction liquid through micro-volume injection, performing centrifugation fusion, and performing amplification on genomes, transcriptome or specific nucleotide sequences at an appropriate temperature. The micro-volume cell nucleic acid amplification method provided by the invention is simple in operation, low in cost and high in flux, and the reaction volume can be reduced to a nano liter grade.

The invention relates to a circulation free DNA and blood cell preservation medium which comprises the following components: 1) one or more protection agents; 2) one or more chelation stabilizers; 3)one or more metabolic inhibitors; 4) one or more enzyme inhibitors; 5) one or more anti-adsorption agents; 6) a buffer solution. The preservation medium has a remarkable circulation free DNA and cellprotection effect, is capable of preserving a blood preparation or a biological sample for a long time at a normal temperature and is free of free aldehyde substance, anaphylactic reactions and toxicreactions of human bodies in contact with liquid samples and biological samples with the preservation medium are reduced, potential carcinogenic risks of the preservation medium are reduced, the completeness of nucleic acid in a product preparation and a biological sample can be maintained, accuracy and scientificity of nucleic acid substance detection can be ensured, and detection results approximate to reality can be provided.

The invention provides a micro-fluidic chip and a nucleic acid extraction and purification method therewith. The micro-fluidic chip has a four-layer membrane structure including: a liquid path layer, a gas path control layer, a magnet control layer and a glass substrate layer. The gas path control layer is arranged below the liquid path layer. The substrate layer is arranged below the gas path control layer. The magnet control layer is arranged below the glass substrate layer. The method, with the micro-fluidic chip, includes the steps of: a) introducing a target sample; b) introducing superpara-magnetic silicon beads for nucleic acid purification; and c) extracting and purifying nucleic acid from the mixed sample. In the invention, introduction of the sample, cleaning of non-nucleic acid substances and collection and purification of the nucleic acid are integrated on one micro-fluidic chip, so that the nucleic acid extraction and purification process is optimized and extraction and purification efficiency is greatly improved. The method simplifies manual operations and achieves temporal-spatial resolution information which is difficult in conventional methods, and has excellent nucleic acid extraction and purification effects.

The invention relates to the technical field of nucleic acid extraction, in particular to a rapid nucleic acid extraction device and a method for extracting nucleic acid. The rapid nucleic acid extraction device comprises a rapid nucleic acid extractor, a lysis solution, a washing solution and an eluent, the eluent is used for eluting and collecting nucleic acid substances adsorbed on the rapid nucleic acid extractor, and the washing solution is one or a mixture of more of silicone oil, paraffin oil, C5-C15 alkane oil and halogenated C1-C15 alkane oil. The washing solution does not contain alcohol, does not need to be aired, does not influence nucleic acid, and ensures quick and effective detection.

To provide a new technique by which efficient transfection is ensured in delivering a target gene into a cell, disclosed is a nucleic acid construct for nuclear import, which comprises a ternary complex consisting of a nucleic acid substance containing a gene to be delivered into the nucleus of a cell, an importin protein (for example, importin-β) capable of passing through the nuclear pore and involved in the nuclear transport, and a binding substance (for example, polyethyleneimine) bound to both of the nucleic acid substance and the importin protein. Nucleic acid transport from outside of a cell into the cell nucleus can be particularly promoted by administering the nucleic acid construct bound to a cell membrane receptor binding factor and/or a membrane fusing substance, or administering the nucleic acid construct encapsulated in a non-viral vector (for example, Sendai virus envelope) having cell membrane permeability and membrane fusing properties.

The invention discloses a method for efficiently extracting high-quality nucleic acid substances from olive leaves. The operation steps mainly include sampling, grinding, pyrolysis, impurity removal,precipitation, purification, dissolution and the like. Aiming at the characteristics that the olive is rich in impurities such as polyphenols, polysaccharides and proteins, on the basis of a traditional CTAB method, deep optimization is carried out on the characteristics of olive leaves, systematic improvement is carried out on sample dosage, reagent types, reagent concentration, reaction time, reaction volume, operation steps and the like, and a method suitable for high-efficiency and high-quality extraction of nucleic acid substances of olive tender leaves, mature leaves or old leaves is developed. The method can select and extract olive leaf genome DNA or total RNA according to actual requirements, has wide applicability and strong practicability, and has good application value.

The invention provides a novel method for extracting phage genome DNA. The novel method comprises the following steps: after amplifying phage lysate, removing cell debris and nucleic acid substances of host bacteria, adding PEG6000 to settle phage, after resuspending by TM buffer liquid, avoiding chloroform extraction, directly adding nuclease, removing the genome of the remaining host bacteria, then replacing protease K by using urea, after degeneration of coat protein, separating the protein from the genome DNA by agarose gel electrophoresis, and then recycling the genome DNA of the phage byusing a freeze thawing and recycling method. The method is simple and convenient to operate, is particularly suitable for phage sensitive to chloroform, and is wide in application range, experiment costs are greatly reduced, and extracting time is shortened.

The invention provides application of Wnt2 to preparation of a medicament for inhibiting the esophageal cancer, and provides a medicament for inhibiting the development of the esophageal cancer. The medicament comprises a component for inhibiting the action of a Wnt2 gene or/and a Wnt2 protein. The component comprises one or more a polypeptide substance capable of enclosing the Wnt2 gene, an antibody/ligand capable of enclosing the Wnt2, a lethal medicament or a nucleic acid substance carried by an antibody/ligand capable of inhibiting or killing the Wnt2 and a small interfering ribonucleic acid (siRNA) or a miRNA (micro Ribonucleic Acid) or a shRNA (short hairpin Ribonucleic Acid) capable of interfering/blocking the Wnt2 gene. The invention has the advantage: the application of the Wnt2 gene to the preparation of a medicament for inhibiting the esophageal cancer is provided, a WNT signal conduction path in a WNT2 activated tumor cell is blocked under the silencing action or the neutralizing action of an antibody, so that the effect of inhibiting the growth of tumors is achieved.

The invention belongs to the technical field of biochemistry, and provides a method for extracting nucleic acid from bee pollen and purifying. The specific method is as follows: freezing screened, purified and dried bee pollen, and ultrafine grinding until the particle size achieves 20-60 meshes, adding a 0.14mol/L sodium chloride solution with weight ratio of 1: (8-10), homogenating, separating organelle through differential centrifugation, and ultrasonically treating for 30-40 minutes at 20-30 KHz, cracking to obtain DNA-proteins, then adding a 1mol/L sodium chloride solution with weight ratio of 1: (6-8), homogenating, ultrasonically treating for 30-40 minutes at 20-30 KHz, cracking to obtain RNA-nucleoproteins, adding a proper amount of sodium dodecyl sulfate to separate the proteins from nucleoproteins, adding concentrated potassium acetate, precipitating a sodium dodecyl sulfate-protein complex, transforming spare sodium dodecyl sulfate as sylvite with small solubility and simultaneously precipitating, repeatedly extracting, obtaining a nucleic acid pure solution through ultracentrifugation or ion-exchange column chromatography, and performing vacuum freezing and drying on the nucleic acid pure solution to obtain the pure nucleic acid substance.

The invention relates to dsRNA in an organism or tissue and a method for extracting the dsRNA. The method comprises the following steps: pre-treating, centrifuging to obtain supernate, carrying out chromatography on the supernate, carrying out segmented elution, and collecting eluent for post-treatment. According to the method, organic reagent treatment and cellulose column chromatography are combined, firstly, an organism or tissue is treated to be broken, and nucleic acid substances are released into a liquid phase; and by means of reversible adsorption of filler in cellulose column chromatography on liquid-phase medium-concentration nucleic acid substances, dsRNA and other nucleic acid substances can be effectively separated through segmented elution, especially interference of DNA degradation substances can be remarkably reduced, and the purity of the final product dsRNA can be improved.

The invention relates to the technical field of biological materials, in particular to a preparation method of decellularized heterogeneous small blood vessels. By adopting a method for combining ultrasound, perfusion, detergent and enzyme, cells in a biological material matrix are broken through a cavitation effect of high-frequency ultrasonic waves, then the detergent is pressed into a materialin a perfusion mode to further wash cell components and part of immunogenic substances; after the primary action of ultrasound and the detergent, the cells are basically broken, most of cell components are washed away, most of nucleic acid substances are further removed under the action of DNase-I and RNase-A, and the residual quantity is less than 100 ng/mg; and finally, a good decellularizationeffect is achieved through repeated sterile PBS solution lavage.

The invention provides a biological detection method based on an excitation light source. The biological detection method comprises the following steps of: injecting a sample into a sample adding holeformed in a sample adding layer; connecting a gasket arranged below the sample adding layer with a pipeline layer at the lower end through a buckle structure, during use, pulling out the gasket alonga rail, pressing the sample adding layer downwards to connect the pipeline layer with the sample adding layer in a clamped mode, sequentially mixing a row of pricking needles arranged on the pipelinelayer with a reagent of the sample adding layer, and carrying out extraction and purification reaction in the pipeline layer, then introducing nucleic acid substances in the sample into an amplification bin arranged on the pipeline layer; enabling an excitation light module to emit exciting light with a preset wavelength to an amplification bin, carrying out reaction, then acquiring fluorescenceinformation through a fluorescence sensor to complete detection, and controlling the temperature in the amplification bin to be within a preset range by the temperature control module, thus enabling areagent of a chip device to reach an optimal reaction state.

The invention provides a eugenol combination liquid as well as a preparation method and an application of the eugenol combination liquid. The combination liquid comprises the following raw materials by volume: 99.8555%-99.9711% of water, 0.01%-0.05% of Tween 80 solution, and 0.0189%-0.0945% of eugenol; and the invention provides the method for preparing the eugenol combination liquid. The eugenol combination liquid inhibits the growth of staphylococcus aureus, and can even be used as a bactericide when the concentration of the eugenol is higher. The eugenol combination liquid affects the molecular structures of an SDH enzyme and an MDH enzyme of staphylococcus aureus, and the staphylococcus aureus is changed, so that the conformation is changed, and the activity of the enzyme is reduced. After the eugenol combination liquid acts on the staphylococcus aureus, the nucleic acid substances related to the synthesis of protein are leaked because of the change of cell permeability, the synthesis of the protein is further inhibited, and finally the cell death is caused.

The invention discloses a magnetic bead mixed liquid split charging device and method for nucleic acid substance extraction. The device comprises a shell, a stirring device is arranged in the shell, the stirring device comprises a stirring part for stirring, the stirring part is arranged at the middle lower part of an inner cavity of the shell, the stirring part is arranged in the shell, the upper end of the stirring part is fixedly connected with the lower end of a suction pipe of a tubular structure, the upper end of the suction pipe is communicated with a negative pressure device through a conveying pipe, a rotating device used for providing rotating force power is arranged on the suction pipe, the negative pressure device and the rotating device are arranged outside the shell, the rotating device drives the stirring part to rotate through the suction pipe, and the suction pipe is used for discharging the magnetic bead mixed liquid in the shell. By means of the split charging device and the split charging method, automatic even mixing and automatic split charging of the magnetic bead mixed liquid can be achieved, the adsorption performance of the magnetic beads can be guaranteed under the low-temperature condition, time and labor are saved, operation is standard, cost is low, and the device and the method are particularly suitable for industrial production.