60results about How to "High reaction specificity" patented technology

The present invention provides reaction mixtures that include blocked oligonucleotides comprising 2′-terminator nucleotides. The blocked oligonucleotides are rendered extendible when the 2′-terminator nucleotides are removed from the oligonucleotides, e.g., via pyrophosphorolysis. The reaction mixtures can be used in various nucleic acid polymerization and / or amplification assays, among many other applications. In addition to reaction mixtures, the invention also provides related methods and reaction mixtures.

The present disclosure provides a method for preparing a radioactive ligand or radioactive substrate having affinity for a target biomacromolecule, the method comprising: (a) reacting a first compound comprising a first functional group capable of participating in a click chemistry reaction, with a radioactive reagent under conditions sufficient to displace the leaving group with a radioactive component of the radioactive reagent to form a first radioactive compound; (b) providing a second compound comprising a second complementary functional group capable of participating in a click chemistry reaction with the first functional group; (c) reacting the first functional group of the first radioactive compound with the complementary functional group of the second compound via a click chemistry reaction to form the radioactive ligand or substrate; and (d) isolating the radioactive ligand or substrate.

A process for producing biological diesel oil by fixed enzyme method is carried out by taking sodium alginate, kaoline, glutaraldehyde, gelatin and lactose as co-fixer, fixing the lipase, piling the spherical fixed enzyme in filled bed columnar reactor naturally, alcoholyzing animal and plant oil fatty with biological diesel oil as solvent and producing biological diesel oil. It is cheap, has less consumption and by product and better quality. It can be used for continuous production.

The present invention provides reaction mixtures that include blocked oligonucleotides comprising 2′-terminator nucleotides. The blocked oligonucleotides are rendered extendible when the 2′-terminator nucleotides are removed from the oligonucleotides, e.g., via pyrophosphorolysis. The reaction mixtures can be used in various nucleic acid polymerization and / or amplification assays, among many other applications. In addition to reaction mixtures, the invention also provides related methods and reaction mixtures.

The present invention is directed to methods of amplification and detection of nucleic acids by rolling circle amplification. The methods of the present invention may be used to amplify nucleic acids for detection and cloning. The methods are particularly suited to RNA.

A system for performing molecular biological diagnosis, analysis and multistep and multiplex reactions utilizes a selfaddressable, selfassembling microelectronic system for actively carrying out controlled reactions in microscopic formats. Preferably, a fluidic system flow a sample across an active area of the biochip, increasing diagnostic efficiency. Preferably, the fluidic system includes a flow cell having a window.

The invention discloses a specific primer probe composition, a kit and a method for detecting a T790M site of an EGFR gene. The kit comprises an upstream primer, a downstream primer, a fluorescent probe I for detecting T790M mutation and a fluorescent probe II for detecting a wild type. For the kit, the primer pair with the specific sequences and the probes with the specific sequences are designedfor the human EGFR gene T790M mutation and reaction systems are optimized. According to the invention, through a method of a Raindropdigital PCR platform, high sensitivity detection on the T790M mutation of the EGFR gene is achieved and the abundance of the mutation is obtained simultaneously; the detection limit can be low to one in 100000. Furthermore, the invention can be used for detecting multi-source samples, comprising tumor tissue samples and ctDNAs, so that the application scope of the kit, reaction systems and method are expanded, and a medication guidance for treatment of patientswith lung cancer T790M mutation is provided.

The method of catalytically synthesizing vitamin A fatty ester with immobilized lipase includes mixing substrate vitamin An acetate and C10-C18 fatty acid or fatty ester in the molar ratio of 1 to 1-7 as well as organic solvent and immobilized lipase in the amount of 0.2-5 times weight of vitamin An acetate for reaction under 20-50 deg.c for 9-50 hr; taking out the immobilized lipase; filtering the reacted liquid and crystallizing to obtain vitamin A fatty ester. The organic solvent is C6-C10 saturated alkane containing water in 0-1 %; and the immobilized lipase is yeast type lipase on carrier of diatomite, resin or fabric membrane. The immobilized lipase of the present invention has the features of great catalysis area, high catalysis efficiency, high stability, capacity of being reused, etc. The reaction solvent has no toxicity, and the product is light in color and high in quality.

The invention relates to a poly alkyl ether compound with a strange end group and double functional groups, which has a formula (I), wherein A is selected from any one of electrophilic group-CONHNH2, -ONH2, or -NH2, and PAG is a poly alkyl ether chain without an end group. The poly alkyl ether compound with a strange end group and double functional groups can directly react with protein, polypeptide or free sulfhydryl and a free carbonyl group of a drug molecule under mild conditions, and is strong in reaction specificity and fast in speed. The poly alkyl ether compound can be used for chemical crosslinking between the protein, the polypeptide and other drug molecules and a carrier to prepare a combination with uniform fixed points.

A system for performing molecular biological diagnosis, analysis and multistep and multiplex reactions utilizes a selfaddressable, selfassembling microelectronic system for actively carrying out controlled reactions in microscopic formats. Preferably, a fluidic system flow a sample across an active area of the biochip, increasing diagnostic efficiency. Preferably, the fluidic system includes a flow cell having a window. Pulsed activation of the electrodes of the biochip are advantageously employed with the fluidic system, permitting more complete sampling of the materials within the biological sample. An improved detection system utilizes a preferably coaxially oriented excitation fiber, such as a fiber optic, disposed within a light guide, such as a liquid light guide. In this way, small geometric systems may be fluorescently imaged. A highly automated DNA diagnostic system results. Perturbation of the fluorescence signal during electronic denaturation is detailed and analyzed for analytical and diagnostic purposes. Such fluorescence perturbation information is combined with other information to provide improved analysis. DNA fingerprinting uses hybridizing DNA fragments of a given length and a capture sequence at a test site and then determining the level of reverse bias necessary to affect the hybridization, such as to dehybridize, and determine the length of the DNA.

The invention discloses a method and a kit for identifying the early embryonic sex of a pig by dual temperature multiplex PCR. A dual temperature multiplex PCR reaction system with rapidness, sensitiveness and high accuracy, which is suitable for identifying the early embryonic sex of the pig, is used for identifying the early embryonic sex of the pig. The kit which is developed by the dual temperature multiplex PCR reaction system and used for identifying the early embryonic sex of the pig fully plays the characteristics of high efficiency, economy, simpleness, convenience and high reaction specificity of the dual temperature multiplex PCR and realizes the rapid identification of the early embryo of the pig, thereby realizing the sex control of the pig, improving the economic benefits ofpig breeding industry, saving the breeding cost and providing convenience.

The invention discloses primers for detecting hepatitis B virus nucleic acids, a probe, a kit and a detection method. A primer pair for detecting hepatitis B virus nucleic acids comprises a first primer and a second primer, and the sequences of the first primer and the second primer are respectively as shown in SEQ ID NO.1 and SEQ ID NO.2. The sequence of the probe is as shown in SEQ ID NO.3. Thekit comprises the primer pair and the probe. The detection method of hepatitis B virus nucleic acids comprises the following steps: mixing a nucleic acid sample to be detected with PCR (Polymerase Chain Reaction) liquid by adopting a sample treatment device, and coating the generated mixed liquid with droplet generation liquid to form droplets which contain single nucleic acids or do not contain nucleic acids; and performing PCR amplification on the droplets by utilizing the kit, and detecting PCR amplification products. The kit disclosed by the invention is low in lower limit of detection, high in sensitivity and good in specificity. The detection method disclosed by the invention is simple and quick, very high in detection efficiency and low in cost.

The invention discloses a rotating packed bed reactor for enzyme catalysis and an application method of the reactor in fatty acid production through enzyme catalysis lipid hydrolysis. The method comprises the steps that lipid and water are adopted as raw materials to be mixed with lipase for a catalysis reaction, the catalysis reaction is conducted under the condition that the supergravity level (gravitational acceleration) is 5 g to 120 g, steel mesh packing is adopted, the substrate flow speed is 80 mL / min to 120 mL / min, the temperature is 35 DEG C to 55 DEG C, the lipase adding amount (lipase / substrates) is 0.5 wt% to 1.5 wt%, and the lipid-water mass ratio is (0.5-1.5):1, free fatty acid is prepared after reacting is conducted for 8 hours to 24 hours, and the hydrolysis yield can reach 98%. According to the method, lipase is used for catalytically producing the fatty acid in the rotating packed bed reactor for the first time, and the advantages that a rotating packed bed is high in mass transfer intensity and mass transfer efficiency, and the materials are full in micromixing in the bed and short in dwell time are achieved in a lipase catalysis technology; in a solvent-free system, compared with a traditional stirring system, the technology has the advantages that the reaction time is shortened by 30%-50%, and the fatty acid yield is increased by 15% to 50%.

The invention belongs to the technical field of biology, and discloses a hybridoma cell strain secreting a porcine sapelovirus VP1 protein monoclonal antibody. The preservation number of the hybridoma cell strain is CCTCC NO: C2021177, and the hybridoma cell strain is preserved in China Center for Type Culture Collection, Wuhan University, Wuhan, China. The hybridoma cell has stable antibody secretion capacity, the secreted monoclonal antibody has good reaction specificity with the porcine sapelovirus VP1 protein, the antigen epitope recognized by the monoclonal antibody is the 40th-46th amino acid of the porcine sapelovirus VP1 protein, the polypeptide sequence of the antigen epitope is 40PALTAAE46, and the epitope is not reported yet. The hybridoma cell strain lays a good material foundation for researching the etiology and pathogenesis of the porcine sapelovirus in the future.

The invention discloses a culture medium for producing benzoylformic acid by fermentation method. The culture medium comprises calcium ion and D, L-phenylglycine and is characterized in that the initial pH value of the culture medium ranges from pH more than 7 to pH equal to 10. By adopting the culture medium to produce the benzoylformic acid, the substrate inventory rating can increase to 50g / L; the conversion rate can reach 36.8%, the response-specific is high, almost no byproduct exists, simple extraction is enough, and the operation is very convenient.

The invention relates to a method for measuring the content of triterpenoids in lucid ganoderma spore oil, which comprises that heating lucid ganoderma spore oil via alcohol in hot water bath at 60-70Deg. C until being dissolved, extracting the triterpenoids in the lucid ganoderma spore oil, to react with vanillin via perchloric acid in glacial acetic acid, using oleanolic acid as reference, using colorimetric method to test the content at 549nm wavelength. The inventive method can check the triterpenoids content of lucid ganoderma spore oil, with strong reaction specificity, stable method, better repeatability, high sensitivity at 0. 0087 / mug, minimum detect limit as 2mug, and label recycle rate as 98. 15%.

The invention discloses an extraction method for mushroom polysaccharide and a preparation method for a double-mushroom soup-stock essence. The preparation method for the double-mushroom soup-stock essence comprises the following steps: (1) uniformly mixing a natural mushroom raw material and a compound enzyme preparation, then adding water, and carrying out heating and activating so as to obtain a mixed solution; (2) adjusting a pH value of the mixed solution to be 5.0, carrying out enzymatic hydrolysis at 40 to 50 DEG C so as to obtain an enzymatic hydrolysate; (3) decocting the enzymatic hydrolysate, carrying out centrifugation, then subjecting an obtained supernatant to filtration, concentration and refiltration, and carrying out precipitation with ethanol; (4) subjecting obtained precipitate to vacuum freezing and drying so as to obtain the mushroom polysaccharide; and (5) with a low-temperature drying technology and a starch-coating forming technology, blending the mushroom polysaccharide and a solid-state compound seasoning product so as to obtain the double-mushroom soup-stock essence. The double-mushroom soup-stock essence provided by the invention has the advantages of flavor-improving and freshness-increasing functions of a common seasoning, nutritional values of health-care products, simple process, less equipment investment, safe and controllable production operation and food quality, stable quality, and applicability to mass production.

The invention belongs to the field of protein detection, and relates to a microfluidic chip colorimetric detection method and kit for detecting creatine kinase isoenzyme. The method comprises the following steps: S1, preparing a linear polyacrylamide-DNA polymer; S2, synthesizing and modifying gold nanoparticles; S3, designing and assembling a micro-fluidic chip; S4, synthesizing DNA hydrogel; and S5, detecting. According to the invention, the DNA hydrogel is used for detecting the target object, the stability is good, the reaction specificity is high, and visual detection can be achieved; and the micro-fluidic chip is used for detection, and the purpose of portable quantitative detection is achieved.

The invention discloses a method for preparing a hot start Taq polymerase. The method comprises the following step of mixing the Taq polymerase with a a treatment solution, so as to obtain the hot start Taq polymerase, wherein the treatment solution is a heparin solution, a heparin sodium solution or a heparin potassium solution. The method for preparing the hot start Taq polymerase has the beneficial effects that the hot start can be realized in the first step of PCR (Polymerase Chain Reaction) reaction during carrying out the PCR reaction, and the hot start also can be realized in the follow-up steps of the PCR reaction, so that the amplification of non-specific sequences can be effectively avoided, and the reaction specificity, reliability, uniformity and sensitivity of the DNA (Deoxyribose Nucleic Acid) polymerase can be improved; compared with the conventional similar product, the hot start Taq polymerase prepared by the method disclosed by the invention has the advantages that the amplification efficiency is increased, the hot start Taq polymerase can be detected from a micro sample and trace sample under relatively low cycle number as well as be cloned to a target gene, thus the personal error can be improved to a great extent; and the hot start Taq polymerase is suitable for being applied to amplification and detection of general PCR reaction, complex templates and trace templates.

The invention discloses a preparation method of a detecting paper tube for cobalt ions, the detecting paper tube and a detecting method. The preparation method comprises the steps that a polyoxyethylene dehydrated sorbitol monooleate solution is added into a nano-gold solution, after stirring for the first moment, a mono(6-mercapto-6-deoxy)-beta-cyclodextrin aqueous solution is added, and a mono(6-mercapto-6-deoxy)-beta-cyclodextrin functionalized gold nanoparticle compound is obtained through stirring; a sample paper pad (5) used for bearing cobalt ion samples, a combining paper pad (6), a detecting paper pad (1) and an absorption paper pad (7) are sequentially wound from the open end (3) to the closed end (4) of a transparent tube body (2) so as to form a winding drum structure; and a color contrast area (9) with the color gradients is arranged on the position, corresponding to the absorption paper pad (7), of the transparent tube body (2), and the different color gradients of the color contrast area (9) correspond to the concentration values of the cobalt ions.

The present invention provides a novel immunoassay method with high reaction specificity and high sensitivity. The present invention also provides a method for immunoassaying a target antigen utilizing reactivation of an apoenzyme, which includes simultaneously or sequentially adding a test sample to an antibody specific to the target antigen, the target antigen labeled with a coenzyme, an apo-D-amino acid oxidase, a D-amino acid, and a reagent for detecting a hydrogen peroxide produced by the oxidase.

The present invention provides a novel immunoassay method with high reaction specificity and high sensitivity. The present invention also provides a method for immunoassaying a target antigen utilizing reactivation of an apoenzyme, which includes simultaneously or sequentially adding a test sample to an antibody specific to the target antigen, the target antigen labeled with a coenzyme, an apo-D-amino acid oxidase, a D-amino acid, and a reagent for detecting a hydrogen peroxide produced by the oxidase.

The invention provides a preparation method of (R)-N-bromine-methyl naltrexone. The preparation method adopting naltrexone as the raw material comprises the following steps of: protecting 3-phenolic hydroxyl by adopting an organic silicon group to obtain a first naltrexone derivative; reacting the first naltrexone derivative with a methylation reagent CH3X to obtain a second naltrexone derivative; and removing the phenolic hydroxyl protective group from the second naltrexone derivative, and carrying out bromine anion exchange while necessary to obtain a target product. The invention further provides the two naltrexone derivatives related in the preparation method. The preparation method provided by the invention is high in directional rate and purity, less in steps, simple to operate, gentle in reaction conditions, low in device requirements and convenient to realize industrial production.

A process for producing biological diesel oil by fixed enzyme method is carried out by taking sodium alginate, kaoline, glutaraldehyde, gelatin and lactose as co-fixer, fixing the lipase, piling the spherical fixed enzyme in filled bed columnar reactor naturally, alcoholyzing animal and plant oil fatty with biological diesel oil as solvent and producing biological diesel oil. It is cheap, has less consumption and by product and better quality. It can be used for continuous production.

The invention discloses a PCR (polymerase chain reaction) specificity improving method. In the method, a sequence which is nonhomologous from a template and incapable of forming stable secondary structure is added to the end 5' of a primer pair; further, reaction specificity is improved by adding tail primer to the end 5' on the basis of compound PCR conditions. The compound PCR conditions include, (1) 5-10 cycles of normal PCR reaction condition and (2) two-step PCR condition based on combination of high-temperature annealing and extension. Compared with normal PCR primer, the tailed primer can improve PCR specificity effectively without influence to flexibility; meanwhile, working concentration of the tailed primer is equal to or even lower than that of common primer, thus, PCR specificity can be improved remarkably on the normal PCR condition, and can be further improved by a way of combining with the compound PCR conditions. The method is adaptable to various PCR such as normal PCR and nested PCR, and is of high application value.

The invention discloses an LSP (linear scorpion primer) and kit for detecting human BRCA1 (breast cancer susceptibility gene 1) mutations, wherein 5' end of the LSP is provided with a linear probe having different dual-fluorescent labels, 3' end is provided with a sequence fragment complementary to a template, and the two sequence fragments are linked via spacer 18; when the template exists, the LSP and the template can be extended by annealing to synthesize a template complementary chain; after the template complementary chain is synthesized, the 5' end probe portion of the LSP may bind with the chain, forming a scorpion-like shape; a downstream primer is extended along the 3' end of the template complementary chain; the probe portion is sheared under the action of a polymerase so as to emit light. The LSP of the invention has good specificity and high sensitivity, approximate genes are free of mutual interference, and detection results show higher fluorescent signal-to-noise ratio.

A clozapine individualized drug gene detection kit comprises the following components: CYP2D6*2 (886C) A, CYP2D6*41 (985+39G) A, CYP2D6*10 (100C) T and MC4R (57882787C) T; 4 pair of site amplificationand sequencing prim, 1 pair of CYP2D6Z large fragment PCR amplification primers, PCR amplification reagent, PCR product purification reagent and DNA sequencing reagent; as shown in Table 1 of that prim sequence. The invention has the following technical effects: a gene detection kit for individualized clozapine medication for Chinese people is provided, and the sensitivity is high and the accuracy is high. The kit is used for CYP2D6*2 (886C) A), CYP2D6*41(985+39G)A), CYP2D6*10 (100C) T) and MC4R (57882787C) T). The genetic difference between individuals can be found through the test of this item, and the drug constitution (slow metabolism or normal metabolism) can be defined, the relevant gene information can be interpreted, and the physiological status or drug reaction can be known in time, so that the drug efficacy can be improved, the toxic and side effects of drugs can be reduced, and the medical cost can be reduced.

The invention discloses a recombinant MSG1 protein monoclonal antibody and a blocked ELISA (Enzyme-linked Immuno Sorbent Assay) method for detecting a haemophilus parasuis mycoplasma specific antibody. A hybridoma cell line 1A7 of an excretion haemophilus parasuis mycoplasma recombinant MSG1 protein monoclonal antibody is preserved with preservation number of CGMCC NO. 4860 and the preservation date of May 13th, 2011. The monoclonal antibody of the haemophilus parasuis mycoplasma recombinant MSG1 protein is excreted by the hybridoma cell line 1A7 with preservation number of CGMCC NO. 4860. The monoclonal antibody as well as the haemophilus parasuis mycoplasma MSG1 recombinant protein and an etiology protein are provided with good reaction specificities. According to the invention, the monoclonal antibody is applied to the blocked ELISA (Enzyme-linked Immuno Sorbent Assay) method for detecting the haemophilus parasuis mycoplasma specific antibody; and the detection condition is optimized through a large quantity of experiments so that the method has the advantages of good repeatability, sensitivity and specificity.

The invention discloses a theileria annulata SYBR Green I real-time fluorescence PCR (Polymerase Chain Reaction) specific primer as well as a detection kit and a real-time fluorescence PCR detecting method. The method is very convenient and quick in detecting, can improve specificity and sensibility of the detecting method, takes trace pathogen genome DNA (deoxyribonucleic acid) extracted from blood or lymph as a template to authenticate, designs two specific primers for a specific area which takes Tams1 as a target gene of theileria annulata, and solves the defects that an existing PCR detecting technology is low in specificity and sensibility, and easily causes leakage detection of false positive and when content of theileria annulata is relatively low during amplification. The method isstronger in reaction specificity, is higher in sensibility, is specially used for precise detection of theileria annulata, and has a wide application prospect.

The present invention discloses a method for optimizing PCR amplification by adding elementary substance material into PCR system, wherein the elementary substance material is selected from a group consisting of element titanium, element nickel, element bismuth, element stibium, element selenium, element chromium, and a mixture of the group. This new method is more effective than conventional amplifying method and could be widely employed in many fronts, especially in multiplex PCR, two-round PCR, low-copy PCR, long-term PCR and rapid PCR.