132results about How to "Small reaction volume" patented technology

The invention discloses a PCR (Polymerase Chain Reaction) chip based on droplet array. The chip is obtained by taking a mono crystalline silicon sheet or a glass sheet as a base material and adopting standard photoetching as well as wet etching technique. The mono crystalline silicon sheet or the glass sheet is the base material of the chip. A SiO2 oxide layer and a silane layer generated by silanization on the surface of the SiO2 oxide layer are adhered to the base material successively. A hydrophilic pit array, which can be a droplet array and is formed by etching with photoresist technology, is disposed on the silane layer. A closed circular guardrail is arranged on the outer periphery of the hydrophilic pit array and the guardrail is connected with the silane layer in a sealed manner. The invention also discloses the application of the chip on real-time quantitative PCR detection and real-time quantitative isothermal amplification reaction detection. The detection system provided in the invention remarkably reduces the reaction volume and the detection sensitivity of the system fairly equals that of a commercialized quantitative PCR instrument. The detection system in the invention is easy for operation and has low cost, thus can be applied to commercialization.

The invention relates to a gene chip for high-flux detection of pathogens and application thereof. The gene comprises (1) a combination of 174 oligonucleotide probes of pathogen variety specific genes, toxin genes and drug-resistant genes; and (2) a probe array, which is formed by curing the oligonucleotide probes on a carrier material by arm molecules. The gene chip comprises 174 gene probes, namely 32 pathogen variety specific gene probes of the following 8 pathogens of Burkholderia mallei, Burkholderia pseudomallei, Brucella, salmonella, Yersinia pestis, Bacillus anthracis, comma bacillus and the like, 25 toxin gene probe of the following 7 toxins of diphtheria toxin, Shiga toxin, staphylococcus enterotoxin, choleratoxin and the like, and 117 drug-resistant gene probes of 17 drug-resistant genes of extended-spectrum beta-lactamase, cephalosporinase, carbapenemase, integrase gene, common gene engineering carrier drug-resistant gene and the like. The gene chip can be used to detect multiple pathogen variety specific genes, toxin genes and drug-resistant genes.

The present invention relates to a high-throughput detection chip for drug resistance gene of bacteria, and an application thereof. The detection chip comprises 117 gene probes, the drug resistance gene probes are selected from 17 categories of drug resistance genes, which comprise extended spectrum beta-lactamase, cephalosporinase, carbapenemase, integrase gene, tetracycline resistance gene, aminoglycoside resistance gene, disinfectant resistance gene, erythromycin resistance gene, macrolide efflux gene, vancomycin resistance gene, multidrug resistance efflux pump gene, mupirocin resistance gene, sulfanilamide resistance gene, tylosin resistance gene, fluoroquinolone resistance gene, chloramphenicol acetyltransferase and commonly-used genetic engineering vector resistance gene. The chip is adopted for detecting the resistance gene of the pathogenic bacteria. The chip is characterized in that: the chip comprises (1) 117-oligonucleotide probe composition and quality control probes of 17 categories of the drug resistance genes; (2) probe arrays, wherein the oligonucleotide probes are solidified on the vector material through arm molecules to form the probe arrays.

The present invention relates to a gene chip for detecting several infections lideases and its application. It can be used for detecting, typing and strain-identifying 7 main infections diseases of epidemic hemorrhagic fever, tsutsugamushi disease, leptospirosis, schistosomiasis, malaria, cholera and hemorrhagic enteritis due to 0157:H7. Said invention can select specific gene probe and PCR primer respectively from S gene of epidemic hemorrhagic fever virus, 56KD protein gene of oriental rickettsia, 23 SRNA gene of leptospirosis, 5D antigen gene of schistosomiasis, SSrRNA gene of malarial parasite, 0157 antigen gene of colibacillus 0157:H7, H7 antigen gene and toxic gene and outer membrane OWP protein gene of cholera vibrio and toxic gene.

The invention relates to the technology of microscale reactor, in particular to a microscale reactor for synthesizing radioactive drug. The microscale reactor is composed of a sealing-in body, a reaction flask, a separation column and an adsorption column, wherein the sealing-in body is sealed by a fluid channel layer, a middle layer and a control channel layer into a whole; the reaction flask, the separation column and the adsorption column are detachably spliced into the sealing-in body. The fluid passage layer comprises a fluid microchannel which causes fluid to communicate with each reaction flask, the separation column and the adsorption column; the control channel layer comprises a control air microchannel and an air chamber corresponding to the fluid microchannel; a microvalve is composed of an air chamber and a middle layer to control the fluid microchannel to open and close. The microfluidic chip of the invention can synthesize common positron nuclide of microscale volume. The invention has simple and easy preparation technology method, needs no expensive equipment, can reduce space and invested capital for protecting equipment for preparing radioactive drug and can reduce the workload of experimenters.

The invention relates to a biological engineering detection product and an application of the gene chip, and in particular relates to a detection gene chip for helicobacter pylori infection individualized treatment. The chip is capable of simultaneously detecting polymorphisms of helicobacter pylori clarithromycin medicine-resistant mutation sites A2142G and A2143G, carbostyril medicine-resistant mutation sites Asn87(N87K) and Asp91(D91G / Y / N), and sites 2C19*2(G6981A) and 2C19*3(G636A) related to metabolism of a proton pump inhibitor on human cytochrome enzyme CYP450. The chip detection method disclosed by the invention is rapid and accurate and is capable of identifying helicobacter pylori etiology, carrying out medicine-resistant analysis of clarithromycin and carbostyril, and analyzing metabolic individual differences of the proton pump inhibitor, so that the detection gene chip is used for guiding individualized administration of helicobacter pylori infection triplex process treatment.

The invention relates to a method for synthesizing Varenicline intermediate 2, 3, 4, 5-tetralin-1, 5-methylene-hydrogen-benzoazepine. The method includes the following steps: under the action of catalyst, mixing amyl nitrite with o-aminobenzoic acid solution to generate diazonium salt, then mixing the diazonium salt with cyclopentadiene, and heating to react to generate compound 1; feeding ozone into the solution of compound 1, after complete conversion, adding reducing agent to generate compound 2, then dripping the compound 2 to the mixed solution of triacetoxy sodium borohydride and benzylamine to generate compound 3 by loop closing; and hydrogenating the compound 3 under the action of palladium and carbon for debenzylation and reduction to obtain M intermediate 2, 3, 4, 5-tetralin-1, 5-methylene-hydrogen-benzoazepine. The method has the advantages of greatly simplifying the method for preparing Varenicline intermediate, being simple in production process and safe in operation, well ensuring no harm to the environment and control on production cost, increasing yield and being capable of becoming a process in great industrial production.

The invention discloses a method for preparing an iodine-125 brachytherapy source core for treating tumor, which comprises the following steps: firstly, putting a standby filamentary silver in a 20-80 percent sodium hypochlorite solution, and performing oscillatory reaction at room temperature for one hour so that the surface of the filamentary silver is covered with a layer of compact AgCl deposition; and secondly, putting the filamentary silver into a mixture solution of an acetic acid buffer solution and a Na<125>I solution to react for one hour so that a layer of even and compact Ag<125>I deposition which is firm and hard to fall off is formed on the surface of the filamentary silver. The reagents used in the method are all conventional ones which are convenient to purchase and have low prices; irritant chlorine is not generated in the process of preparation; the surface of the iodine-125 brachytherapy source core is even and compact; the utilization rate of the radioiodine is high; and the process time is short, thus mass production is easy to realize.

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 discloses a method for producing miconazole nitrate on an industrialized basis. The method comprises the following steps: A, conducting the Friedel-crafts reaction between chloroacetic chloride and m-dichlorobenzene to prepare 2-chloro-1-(2,4-dichlorophenyl)ethanone; B, conducting the N-alkylation reaction to prepare 1-(2,4-dichlorophenyl)-2-(1H-imidazolyl)ethanone; C, conducting the potassium borohydride reduction reaction to prepare 1-(2,4-dichlorophenyl)-2-(1H-imidazolyl)ethylate; and D, conducting the O-alkylation reaction to prepare the miconazole nitrate as the target product. The method of the invention has the overall advantages of industrialized application value, low cost, easily accessible materials, mild reaction conditions and easy operation; the total yield is not lower than 64%; and the purity of the miconazole nitrate product is higher than 99%.

The invention discloses a bispecific antibody biological activity and titer detection method. The method includes: detecting two antigen binding sites of a bispecific antibody at the same time; performing four-parameter fitting according to a fluorescence ratio and the concentration of a standard sample to obtain a standard curve; calculating the concentration of a to-be-tested sample according to the standard curve. By adoption of the method, intact antibody modules and cell strains high in bispecific binding activity and expression can be screened out more effectively; the method has the advantage of low reaction volume, and experiment reagent cost can be saved maximally; the method can be widely popularized and applied to CLD clone high-throughput screening.

The invention discloses a method for continuously producing atorvastatin calcium by using a micro-reaction device. The production method provided by the invention is simple in process, can realize continuous production, and has relatively high operation safety and selectivity; in addition, since a reaction volume is small and reaction time is short, corrosion to equipment is small; and meanwhile,as a micro-channel reactor has the characteristics of efficient heat and mass transfer capacity and easiness in direct amplification, a reaction conversion rate is high, the obtained atorvastatin calcium is good in quality and low in energy consumption, and the method is an environment-friendly and efficient method for synthesizing atorvastatin calcium.

The invention discloses a biosensor achieving AChE oriented immobilization and a preparation method and application of the biosensor. The preparation method of the biosensor comprises the following steps of preparation of a magnetic nano material MNP, modification, MNPs-AChE preparation and sensor construction. The biosensor is used for detecting residual organophosphorus pesticide. According to the biosensor achieving AChE oriented immobilization and the preparation method and application of the biosensor, acetylcholinesterase is covalently immobilized to the surfaces of terminated carboxyl-modified MNP granules, and an obtained composite nano material MNPs-AChE can maintain enzymic activity to the maximum extent; according to the constructed amperometric biosensor, an MNPs-AChE modified SPCE system can be automatically adjusted through an external magnetic field, electrode modification can be achieved only through one step, and the stability and activity of AChE are improved; good sensitivity and repeatability are shown in the process of detecting an organophosphorus reagent, and the reaction volume is small; the method for preparing the biosensor is easy and convenient to operate, preparation is convenient, the cost is low, and the method is suitable for industrial large-scale production.

The invention relates to a biological engineering detection product and an application of the gene chip, and in particular relates to a detection gene chip for helicobacter pylori infection individualized treatment. The chip is capable of simultaneously detecting polymorphisms of helicobacter pylori clarithromycin medicine-resistant mutation sites A2142G and A2143G, carbostyril medicine-resistant mutation sites Asn87(N87K) and Asp91(D91G / Y / N), and sites 2C19*2(G6981A) and 2C19*3(G636A) related to metabolism of a proton pump inhibitor on human cytochrome enzyme CYP450. The chip detection method disclosed by the invention is rapid and accurate and is capable of identifying helicobacter pylori etiology, carrying out medicine-resistant analysis of clarithromycin and carbostyril, and analyzing metabolic individual differences of the proton pump inhibitor, so that the detection gene chip is used for guiding individualized administration of helicobacter pylori infection triplex process treatment.

The invention relates to the field of microfluidic chips, in particular to a microfluidic chip, a device containing the microfluidic chip and a method for detecting or sorting samples. According to the microfluidic chip for detecting and / or sorting caught single cells provided by the invention, unicellular microemulsion liquid drops can be controlled to enter a cap-shaped chamber 3, so that each cap-shaped chamber 3 only contains one microemulsion liquid drop, a dual-purpose electrode 6 for detecting / catching is arranged in the cap-shaped chamber 3 or a cavity, whether the microemulsion liquiddrops in the cap-shaped chambers 3 contain cells or not can be detected, and thus sorted catching or a reaction can be performed.

The invention relates to a preparation method of high-purity biapenem, which includes the steps of: (A) carrying out a condensation reaction with a compound represented as the formula V and a compound represented as the formula VI in an acetonitrile solvent in the presence of a less amount of N,N-dimethylformamide and an organic alkali to obtain a compound represented as the formula VII; (B) carrying out a catalytic hydrogenation to the reaction product in the step (A) in a mixed solution composed of an alcohol solvent and a non-proton polarity organic solvent in the presence of a catalyst and an organic alkali to remove a carboxylic acid protective group in the compound represented as the formula VII; and (C) filtering a reaction mixed solution to obtain a filter cake, adding the filter cake in water with stirring to obtain a filtrate, mixing the filtrates, adding an organic solvent under stirring, performing precipitation crystallization, and filtering and drying a product to obtain the biapenem.

The invention relates to the field of biotechnology, and discloses a deoxyribonucleic acid (DNA) probe for detecting cryptococus neoformans. The DNA probe for detecting the cryptococus neoformans has a nucleotide sequence shown as SEQ ID No. 1 in a sequence table, and has high specificity and sensitivity of 0.04ng for detecting the cryptococus neoformans. The invention also provides a gene chip of the DNA probe. By using the gene chip, high-flux, automation and quick detection of a sample which possibly contains the cryptococus neoformans can be realized, the specificity is high, the accuracy of a detection result can be guaranteed, and the DNA probe and the gene chip have broad application prospect.

The invention provides a method for regulating a ZSM-5 molecular sieve by using phosphorus species through aerosol auxiliary synthesis. According to the method, an aluminum source, a silicon source, water and a phosphorus source are mixed; an obtained mixture is uniformly stirred to form mixed liquid; aerosol is formed through an aerosol device; drying is performed to obtain amorphous form silicon aluminum phosphorus oxide; the silicon aluminum phosphorus oxide and a tetrapropylammonium hydroxide water solution are mixed; sealing is performed; crystallization is performed at 100 to 200 DEG C, wherein the crystallization time is 3h to 240h; obtained solid products are washed and dried or are directly dried to be roasted; the ZSM-5 molecular sieve is obtained. By the method, the precise regulation on the ZSM-5 molecular sieve morphology can be realized. The method has the advantages that the regulation process is simple; the template agent consumption is low; the pollution is small; the yield is high; good industrial application prospects are realized.

The invention discloses a 17 alpha-acetoxy-6-methylenepregn-4-ene-3,20-dione preparation method, which comprises: carrying out an etherification reaction, a Mannich reaction and an elimination reaction on 17 alpha-hydroxyprogesterone acetate represented by a formula (I) and used as an initial raw material in a tubular reactor to obtain a product 17 alpha-acetoxy-6-methylenepregn-4-ene-3,20-dione represented by a formula (II), wherein the reaction formula is defined in the specification. According to the present invention, by using the tubular reaction mode approximate to plug flow, the materials are hardly backmixed during the reaction, the mass transfer and heat transfer efficiency is good, the generation of side reactions is remarkably reduced, and the method has advantages of high product yield, high product purity, easy operation, few three-waste and the like, and is suitable for industrial production.

Belonging to the field of gene chips, the invention relates to a gene chip for detection of cerebrospinal fluid pathogens. The gene chip provided by the invention includes a substrate and probes, and the probes are fixed on the substrate. Specifically, the probes are nucleotide sequences as shown in SEQ ID NO.1-153, and the substrate is one of a glass slide, a silicon chip, a film and a high polymer material. The gene chip provided by the invention has the advantages of simple operation step, high detection specificity, good stability, short time and low cost, and can realize rapid, low cost, large flux and automatic detection of the species of pathogenic bacteria in cerebrospinal fluid samples of purulent meningitis patients, thus providing help for diagnosis of purulent meningitis.

The invention provides a DNA probe, a reagent kit and a method for detecting deoxyribonuclease. The hairpin-shaped DNA probe serves as a substrate of deoxyribonuclease, and a fluorescence group and a corresponding fluorescence quenching group are marked at the two ends or the interior of the probe respectively. When the structure of the probe is complete, the fluorescence group and the fluorescence quenching group are located on the same DNA molecule, excited fluorescence of the fluorescence group is absorbed by the fluorescence quenching group, and the fluorescence group does not emit fluorescence; when the probe is degraded by deoxyribonuclease, the fluorescence group and the fluorescence quenching group are separated, and the fluorescence group can emit fluorescence normally. Whether deoxyribonuclease exists in a reaction system or not can be judged as long as the intensity of fluorescence emitted from the reaction system is detected with a real-time quantitative PCR instrument. The method can be widely applied to quality detection on residues of deoxyribonuclease in biological products or quantitative detection of deoxyribonuclease.

The invention discloses a lipase mutant and an application thereof. According to the lipase mutant and the application thereof, wherein the lipase mutant has a sequence with amino acid mutation of a sequence shown in SEQ ID NO: 1, and sites with amino acid mutation comprise a V154L site. The lipase mutant provided by the invention realizes the change of protein structure and function, the solubility of the lipase mutant expressed in escherichia coli is obviously improved, and the catalytic activity of enzyme is greatly improved. And in practical application, the enzyme dosage is greatly reduced, the reaction volume is reduced, the post-treatment difficulty is reduced, and the lipase mutant is suitable for industrial production.

The invention relates to an ester exchange reaction method and a device thereof in the production of biodiesel. The steps are as follows: conveying oil, methanol and catalyst solution to an ester exchange raw material mixing tank for pre-esterification; after animal fat and vegetable oil going through the pre-esterification leave the raw material mixing tank and pass through a static mixer, continuously conveying the animal fat and the vegetable oil by an ester exchange reaction charge pump to enter an ester exchange combined reactor through a raw material preheater; and finally leading the materials in a continuous reactor for heat insulation ester exchange reaction. Compared with the prior art, the esterification product does not need to be neutralized and washed, thus reducing the discharge of sewage; continuous production is available, the production efficiency is high and the conversion rate of glyceride reaches 98.5 percent; and by adopting the continuous reactor in the device, the reaction volume is greatly decreased and the production investment is lowered.

The invention belongs to the field of organic synthesis, and particularly relates to a continuous nitrification method of a naphthalene sulfonate compound. The method comprises the steps of simultaneously and continuously making the nitrified matter naphthalene sulfonate compound and a nitrating agent enter a channel of a stainless steel annular reaction device under a water bath and supersonic environment through a metering pump on the basis that the nitrate ratio is 1.0-1.4 or 2.1-2.4, performing a mixing reaction for 40-120 s under mechanical action generated by supersonic and at 20-70 DEG C, and conducting discharging continuously after the reaction. According to the continuous nitrification method of the naphthalene sulfonate compound, a sulfuric acid solution and acid for nitration of the naphthalene sulfonate compound enter the annular channel simultaneously and continuously through the metering pump, and are mixed sufficiently by suffering from influences of mechanical effects of vibration, internal friction and the like generated by centripetal force and supersonic of liquid flow exerted on a liquid, and the reaction is completed.

The invention discloses a method for preparing epoxy propane by using a micro reaction device. The method comprises the following steps: (1) mixing a hydrogen peroxide solution, a formic acid solution and a sulfuric acid solution so as to obtain a homogeneous solution; (2) introducing nitrogen into the micro reaction device; (3) when pumping the homogeneous solution obtained in the step (1) into the micro reaction device, switching the nitrogen introduced into the micro reaction device into a propylene gas, performing reaction, after the reaction is completed, collecting an effluent, thereby obtaining the epoxy propane. The method is rapid in reaction, high in yield, low in cost, high in security, good in operability, small in reaction size, short in time, relatively small in equipment corrosion, simple in production method process, capable of realizing continuous production, relatively high in operation security and relatively high in selectivity, can be directly amplified and is beneficial to industrial production.

The invention discloses a method for catalyzing propylene epoxidation in a micro-channel by utilizing a metalloporphyrin loaded titanium silicate molecular sieve. According to the method, metalloporphyrin with catalytic activity on epoxidation reaction is grafted to a modified titanium silicate molecular sieve through a grafting method to prepare a novel catalyst with higher activity, and furthermore, the propylene epoxidation is catalyzed in a micro-reactor; a micro-channel reactor has micron-grade dispersion scales and continuous flowing of two phases, and fluid can obtain an extremely largespecific surface area, so that the fluid is subjected to efficient heat exchange with a wall surface and more deal mass transfer and heat exchange efficiency is obtained; compared with a traditionalreactor, accurate temperature, pressure and reaction time in the micro-reactor are controlled; and obtained propylene oxide has good selectivity, high yield and great safety and meets the industrial requirements better.

The invention relates to a virus sample treating fluid and treating method of novel coronavirus and a rapid constant-temperature reverse transcription amplification kit for detecting the novel coronavirus, and belongs to the technical field of pathogenic microorganism in-vitro molecular detection. The virus sample treating fluid of the novel coronavirus is a guanidine hydrochloride aqueous solution with the final concentration of 0.1-1 M or a 0.1xTE buffer solution containing Tween-20 with the final mass percentage of 0.01%-0.1%. A virus sample treated with the virus treating fluid can realizesimple, convenient and rapid virus nucleic acid detection, and a result can be observed only by constant-temperature amplification and color development methods.

The invention discloses automatic reaction device and method for pretreating cells. The device comprises a reaction box body, a base, a box cover and a piston, wherein a middle cavity is formed in the center of the reaction box body; seven cavities are formed in the periphery of the hollow cavity; seven square cavities are correspondingly formed between the middle cavity and the seven cavities in the periphery; a control valve is put into each square cavity; communication between an inner chamber and an outer chamber is controlled through up and down movement; the lower part of the reaction box body is tightly connected with the base; the box cover is arranged at the upper part of the reaction box body; a piston rod which is matched with the middle cavity penetrates through the box cover; and the outer side of a second PCR reagent cavity is connected with a PCR chip for detecting nucleic acid. According to the reaction device and method, crushing and cracking of original samples, and purifying and extracting of the nucleic acid and PCR amplification are integrated in a reaction box; the operation process is simplified; the pretreatment efficiency is greatly improved; meanwhile, the reaction volume is reduced; the portability of an instrument is improved; and a foundation is laid for rapid, safe and accurate nucleic acid detection.

The invention discloses a method for synthesizing substitution N-phenylmorpholine compounds. Substituted aniline and 2-chloroethyl ether are heated under the effect of alkali and then are subjected to ring closure to obtain the N-phenylmorpholine compounds. The method has the inexpensive advantages that the inexpensive aniline and the excessive2-chloroethyl ether are directly heated under the effect of the alkali and then are subjected to ring closure to obtain products, reaction is carried out without solvents, accordingly, N-phenylmorpholine of phenylmorpholine with steric hindrance can be synthesized from the aniline steric hindrance by the aid of simple and effective processes, and the cost can be lowered to a great extent; the organic alkali is used, and the method is high in yield.