288 results about "Polymerase chain reaction" patented technology

The invention discloses a molecular marker closely linked with a wheat powdery mildew resistance gene PmCWI16926. The molecular marker is YTU2BS-P12, and the molecular marker is a molecular marker PmCWI16926. The nucleotide sequence of an upstream primer of the molecular marker is as shown in SEQ ID NO: 1; the nucleotide sequence of the downstream primer is as shown in SEQ ID NO: 2; the molecular marker is used for carrying out PCR (Polymerase Chain Reaction) amplification on to-be-detected wheat genome DNA (Deoxyribose Nucleic Acid) to obtain a corresponding amplification product with the molecular weight According to the molecular marker YTU2BS-P12 closely linked with the wheat powdery mildew resistance gene PmCWI16926, provided by the invention, a genetic mapping group of the PmCWI16926 can be more efficiently detected, and map-based cloning of the PmCWI16926 is facilitated; when the marker is used for molecular marker-assisted selection of PmCWI16926, the breeding period can be shortened, the breeding efficiency can be improved, and the marker can be better applied to wheat breeding for disease resistance.

The invention discloses a variable region amino acid sequence of a monoclonal antibody of a West Nile virus non-structural protein NS1 and application of the variable region amino acid sequence, and belongs to the technical field of medicines. According to the invention, West Nile virus non-structural protein NS1 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the West Nile virus non-structural protein NS1 are screened from rabbit spleen cells through flow sorting, and a signal peptide and a variable region gene fragment of an antibody are cloned through reverse transcription-polymerase chain reaction; according to the present invention, the non-structural protein NS1 of flaviviridae flaviviridae virus is taken as a template, and is connected with a constant region gene to an expression vector, and after mammalian cell expression and purification, the monoclonal antibody which has high affinity and is not combined with the non-structural protein NS1 of other eight viruses of flaviviridae flaviviridae virus is obtained through enzyme-linked immunosorbent assay; the monoclonal antibody has application value in diagnosis and prevention and treatment of West Nile virus infection.

The invention belongs to the technical field of genetic engineering, and relates to the field of wheat genetic engineering, in particular to a gene TaABCG42 for regulating and controlling cadmium accumulation of wheat as well as an encoding protein and application of the gene TaABCG42. According to the invention, a TaABCG42 gene is knocked out by using CRISPR / Cas9, a TaABCG42 gene overexpression plant is constructed at the same time, and a homozygous mutant is obtained by using a Hi-Tom sequencing technology and PCR (Polymerase Chain Reaction) successive generation screening. A wild type TaABCG42 homozygous overexpression and knockout plant treated with a 40 [mu] mol / L Cd solution for 5 days is subjected to analysis such as metal content determination, subcellular component metal determination, Cd chemical form determination and Cd ion flow determination, and the Cd absorption and transport approach of TaABCG42 is analyzed, so that the physiological and molecular mechanism of the TaABCG42 metal transporter for absorbing and transporting Cd is explored, and the application prospect is wide. The understanding on the wheat ABC metal transporter is enriched, and a theoretical basis is provided for cultivating high-quality wheat with low Cd accumulation.

The invention relates to the technical field of pathogen detection, in particular to a primer probe combination and a kit for detecting various invasive fungi and application of the primer probe combination and the kit. According to the primer and probe combination provided by the invention, a digital PCR (Polymerase Chain Reaction) platform technology is utilized, so that the rapid, efficient, accurate, sensitive and high-specificity simultaneous quantitative detection of Aspergillus fumigatus, Aspergillus, Mucoriums, Cryptococcus and Pneumocystis jirovecii is realized. The rapid detection of the common invasive fungi in sputum, alveolar lavage fluid, urine, blood and other samples is realized, the detection process does not need culture, the detection period is short, the manpower is greatly reduced, and the detection efficiency is improved.

The invention discloses a primer and a method for quantitatively monitoring the biomass of sargassum hemiphyllum based on an environmental DNA technology, and belongs to the technical field of molecular ecology. The primer probe group comprises an upstream primer, a downstream primer and a fluorescent probe which are specifically targeted to the sargassum hemiphyllum mitochondria COX1 gene, and the sequence is shown as SEQ ID NO.1-3. The kit comprises the primer probe group, a qPCR (quantitative polymerase chain reaction) premixed solution, nuclease-free water and a sargassum hemiphyllum plasmid positive control. The method comprises the following steps: collecting a water sample, enriching eDNA, performing qPCR detection by using the primer probe group after extraction and purification, and realizing qualitative detection and quantitative evaluation of sargassum hemiphyllum through a Cq value or a standard curve. The method disclosed by the invention has the advantages of high sensitivity, strong specificity, no damage to the environment and target organisms, capability of realizing large-scale rapid general survey and the like, and is suitable for early warning of gulfweed blooms, investigation of population distribution and evaluation of ecological influence.

The invention relates to the technical field of bioengineering, in particular to a penicillin G acylase mutant, polynucleotide, an expression vector and application. The penicillin G acylase mutant is obtained by carrying out site-directed mutagenesis on a wild type penicillin G acylase gene of parent Escherichia coli, and carrying out site-directed mutagenesis on the wild type penicillin G acylase gene to obtain the penicillin G acylase mutant. Phenylalanine (F) at the 24th site of an alpha chain, proline (P) at the 383rd site of a beta chain, threonine (T) at the 384th site of the beta chain, glutamic acid (G) at the 385th site of the beta chain and serine (S) at the 386th site of the beta chain of the penicillin G acylase are introduced and mutated by a whole plasmid PCR (Polymerase Chain Reaction) technology to obtain mutants. When the penicillin G acylase mutant obtained by the invention is used for catalyzing methyl mandelate and 3-(1-methyl-1H-tetrazole-5-yl) thiomethyl-7-aminocephalosporanic acid to synthesize cefamandole, the synthesis activity is improved, and meanwhile, the side reaction rate is greatly reduced.

The invention discloses a protein quantitative detection method which comprises the following steps: respectively constructing a capture probe for capturing antibody coupling magnetic beads and oligonucleotides and a detection probe for detecting antibody coupling oligonucleotides through a biorthogonal chemical coupling method; then, target protein is captured by using a biorthogonal chemical coupling and magnetic bead sandwich method comprising the steps of capture probe combination, closing, washing, detection probe combination and the like, and accurate quantification of low-abundance protein in a complex sample is realized by combining a subsequent digital PCR (Polymerase Chain Reaction) technology.

The embodiment of the invention discloses a preparation method of a kit applied to endometrial cancer microsatellite state detection. The preparation method comprises the following steps: taking a first preset volume of polymerase chain reaction amplification liquid; taking a polymerase chain reaction primer mixed solution with a second preset volume; and combining the weighed polymerase chain reaction amplification liquid and polymerase chain reaction primer mixed liquid into the kit applied to endometrial cancer microsatellite state detection. According to the embodiment, the accuracy and the stability of microsatellite state detection of the endometrial cancer can be improved, so that the kit consumed by repeatedly detecting the microsatellite state in the endometrial cancer is reduced.

The PCR plate according to the present invention includes: a body portion having one or more reaction wells; an insertion portion extending from the body portion, inserted into a nucleic acid extraction cartridge, and having an injection port into which a nucleic acid solution is injected; a flow path portion allowing the nucleic acid solution to flow from the injection port to the reaction wells; and a blocking portion attached to the body portion for blocking backflow of the nucleic acid solution from the reaction wells to the flow path portion side.

The invention provides a computer-implemented method and system for non-invasive and step-by-step prediction of cancer risks and a storage medium. The method comprises the following steps: (1) determining a cancer signal score based on the level of a biomarker in a blood sample of a subject; (2) comparing the cancer signal score with a preset threshold value, and determining a primary screening positive subject; (3) further performing cancer risk determination and cancer species prediction by adopting an NGS method on the basis of the primarily screened positive subjects; wherein the biomarker is selected from at least one of AFP (Alpha Fetal Protein), CA125, CA15-3, CA19-9, CA72-4, CEA (Carcino Embryonic Acid), CYFRA (Cytochrome Factor Receptor Amplification) 21-1, ProGRP (Proso-Growth Radical Polymerase Chain Reaction), SCCA (Sequence

The utility model discloses a PCR (Polymerase Chain Reaction) eight-joint pipe cover opening and closing device which comprises a bottom plate, a lower comb plate, a vertical plate, a linear guide rail, an upper comb plate mounting plate, an upper comb plate and a quick clamping device, the lower comb plate and the vertical plate are mounted on the bottom plate in a front-back manner, and a plurality of holes for placing and fixing PCR eight-joint pipe bodies are formed in the lower comb plate; the linear guide rail is vertically arranged on the vertical plate, the upper comb plate installation plate is movably arranged on the linear guide rail through a sliding block, the upper comb plate is installed on the upper comb plate installation plate and corresponds to the lower comb plate in an up-down mode, and the upper comb plate is provided with an open groove capable of clamping a PCR eight-connection pipe cover. The rapid clamping device is installed on the vertical plate through a rapid clamping device installation plate, a handle is arranged on the upper portion of the rapid clamping device, a transmission shaft is arranged on the lower portion of the rapid clamping device, and the transmission shaft of the rapid clamping device is connected with the upper comb plate installation plate through an adjusting nut. The reaction kettle is convenient to use, time-saving and labor-saving, and can avoid the problems that the reaction liquid is polluted and the tube cover is not tightly pressed.

A fiber optic device and method for uniform illumination of biological samples distributed over one or more microcell arrays of a microwell plate can establish more reliable polymerase chain reaction (PCR) data sampling. In some embodiments, the apparatus includes a light engine that emits a light beam captured by a fiber optic bundle for delivering one or more light beams to each microchamber; each light beam thereby fluoresces the biological sample. The fiber optic cable of the fiber optic bundle comprises a distal output end held in place using a mechanical support for flexibly aligning the output beam of the distal output end to be arranged such that the center of the output beam is directed to different locations in the one or more microcell arrays, wherein each of adjacent ones of the output beams partially overlaps so that the light distribution is uniform.

The present disclosure provides an application method in rice callus differentiation based on an Oryza sativa Leafy Cotyledon 1 (OsLEC1) gene, including the steps of: selecting guide ribonucleic acid (gRNA) target sites; cloning tandem fragments including gRNA; ligating each gRNA fragment; performing polymerase chain reaction (PCR) amplification on a ligation product; performing enzyme digestion on the purified product and a target vector; transforming the ligated vector; performing Agrobacterium-mediated genetic transformation of rice; and screening and identifying transgenetic plants. In the present disclosure, the differentiation of callus directly affects the emergence efficiency of transgenic plants. The knockout of OsLEC1 can promote the differentiation of rice callus, suggesting that OsLEC1 can serve as an important target gene for improving the transformation efficiency of rice and even gramineous crops. OsLEC1 can be used as a starting point to construct various molecular tools to enhance transformation efficiency.

The invention discloses a scratch-resistant PCR-PP (Polymerase Chain Reaction-Polypropylene) material and a preparation method thereof. According to the material, through a synergistic modification method of surfactant coupling and chemical crosslinking modification, the recycled polypropylene PCR is combined with a PP matrix, and the scratch resistance, the mechanical property and the processability of the composite material are remarkably improved. The scratch-resistant PCR-PP material comprises a modified PCR material, polypropylene, a micromolecular organic modifier, an antioxidant, a heat stabilizer, a light stabilizer and a filler, and can effectively improve the compatibility of recycled polypropylene and a polypropylene matrix and enhance the surface hardness of the recycled polypropylene and the polypropylene matrix. The material is suitable for high-frequency friction and high-temperature environments, has a wide application prospect, is particularly suitable for the fields of automobiles, electronics and electric appliances, and has good economic benefits and environmental benefits.

The application relates to the field of biological experimental consumables, and discloses an evaporation-preventing reaction plate device for PCR (Polymerase Chain Reaction) reaction. In the application, the PCR reaction plate body is provided with a top cover layer, a heating layer is fixedly arranged in the middle of the top cover layer, a heat cover is fixedly arranged at the top of the heating layer, and a silica gel sealing ring is arranged at the periphery of the bottom of the heat cover. A heating device is arranged at the top of the PCR reaction device, the heating device heats the heat cover to about 105 DEG C, the temperature of the heat cover is higher than that of the reaction liquid, the reaction liquid is effectively prevented from evaporating, the sealing property of the reaction tube is further improved through the silica gel sealing ring at the bottom of the heat cover, each heat cover can be used independently, the heat cover can be directly taken out by buckling from the top, a dust shield is arranged at the top to effectively intercept dust, the reaction device is convenient to use normally next time, the reaction plate at the bottom and the reaction tube groove are integrally quickly detachable and installable, and are convenient to clean and replace.

Clinical management of human cancer is dependent on the accurate monitoring of residual and recurrent tumors. We have developed a method, called personalized analysis of rearranged ends (PARE), which can identify translocations in solid tumors. Analysis of four colorectal and two breast cancers revealed an average of nine rearranged sequences (range 4 to 15) per tumor. Polymerase chain reaction with primers spanning the breakpoints were able to detect mutant DNA molecules present at levels lower than 0.001% and readily identified mutated circulating DNA in patient plasma samples. This approach provides an exquisitely sensitive and broadly applicable approach for the development of personalized biomarkers to enhance the clinical management of cancer patients.

Systems and methods for the detection of sepsis and treatment of patients. A sepsis detection system includes a first subsystem configured to detect the presence of an infection in a patient, a second subsystem configured to detect the presence of a dysregulated immune response in the patient, a third subsystem configured to detect organ dysfunction in the patient, a fourth subsystem configured to detect antibiotic resistance (ARB) of a pathogen in the patient, and a processing device. The first, second, third, and fourth subsystems and the processing device are communicatively coupled to each other via a network. The processing device is configured to determine the presence of sepsis in the patient based on the presence of infection, the presence of the dysregulated host response, and clinical data indicative of organ dysfunction in the patient.The subsystems utilize at least one polymerase chain reaction (PCR) procedure, Raman spectroscopy, clinical data, electronic health record (EHR) data, and antibiotic susceptibility testing (antibiogram).

Provided is an accurate guide RNA (gRNA) screening method for base editing of an asialoglycoprotein receptor 1 (ASGR1) gene, including the following steps: (1) gRNA design; (2) primer design; (3) in vitro transcription of gRNA; (4) cell transfection; (5) collection of cells, and extraction and polymerase chain reaction (PCR) of a genome; and (6) Sanger sequencing.

Systems and methods that enable analyte detection in a multiplexed amplification process can include obtaining, at multiple time points during the amplification process, composite fluorescence signal data associated with a composite fluorescence signal from at least a first probe type comprising a first fluorophore and a second probe type comprising a second fluorophore which has substantially overlapping spectral characteristics as said first fluorophore, the first probe type and the second probe type differing in thermal and / or temporal properties; and determining, based at least partially on the composite fluorescence signal data, fluorescence signal data associated with a fluorescence signal from a given probe type of the first probe type or the second probe type during the amplification process.

The invention discloses a preeclampsia biomarker and an application of the preeclampsia biomarker. The preeclampsia biomarker comprises at least one of a fusion gene PSPC1-MRPS31P2 and a fusion gene LINC00630-AL035494. The preeclampsia biomarker can be used for detecting preeclampsia. The preeclampsia biomarker provided by the invention can provide a non-invasive diagnosis scheme for clinical prenatal preeclampsia, and preeclampsia specific high-expression fusion genes (PSPC1-MRPS31P2 and LINC00630-AL035494) in peripheral blood of a pregnant woman are detected through real-time quantitative PCR (Polymerase Chain Reaction), so that preeclampsia early prediction, disease diagnosis and monitoring of the state of a patient in the illness period are realized; and in combination with the existing clinical diagnosis scheme, the accuracy of pre-eclampsia diagnosis is improved.

The invention discloses a yellow fever virus envelope protein monoclonal antibody and application thereof, and belongs to the technical field of medicines. Yellow fever virus envelope protein E expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the yellow fever virus envelope protein E are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. According to the present invention, the monoclonal antibody with high neutralizing activity and capable of completely protecting mice from yellow fever virus lethal attack is obtained by using the monoclonal antibody as a template, connecting the monoclonal antibody and the constant region gene to an expression vector, carrying out mammalian cell expression and purification, and carrying out enzyme-linked immunosorbent assay, in-vitro virus neutralization and mouse virus attack experiment, such that the monoclonal antibody has high neutralizing activity and can completely protect mice from yellow fever virus lethal attack; the monoclonal antibody has application value in prevention and treatment of yellow fever.

An apparatus for amplifying segments of nucleic acid in a mixture sample via polymerase chain reaction (PCR) has a body including a heating element and a Mixture sample conduit. The heating element includes a denaturation zone, an extension zone and an annealing zone arranged on the body The Mixture sample conduit includes an inlet, an outlet and an intermediate section that passes serially through the denaturation zone, the annealing zone and the extension zone for multiple cycles. A related method for amplifying segments of nucleic acid in a mixture sample via polymerase chain reaction is also described.

The utility model relates to a sample adding device for PCR (Polymerase Chain Reaction), which comprises a base, a containing groove with an upward opening is arranged on the base, a plurality of placing parts are arranged in the containing groove, and a plurality of test tubes can be placed on each placing part. The placing part comprises a first placing plate, a second placing plate and a connecting plate for connecting the first placing plate and the second placing plate, the first placing plate is used for fixing the upper part of a test tube, the second placing plate is used for supporting the bottom of the test tube, and each first placing plate is provided with a connecting part for locking the first placing plate and the adjacent first placing plate. According to the sample adding device for PCR, the connection relation between the two adjacent placing parts can be adjusted according to the row number of test tubes needing to be taken out for observation, so that the efficiency is improved.

The invention relates to the technical field of plant pathogen identification, and provides primers and a method for rapidly detecting and identifying pathotypes of plasmodiophora brassicae based on a PCR (Polymerase Chain Reaction) technology. Comprising an upstream primer and a downstream primer, the nucleotide sequence of the upstream primer is shown as SEQ ID NO. 1 and SEQ ID NO. 2, and the nucleotide sequence of the downstream primer is shown as SEQ ID NO. 3; any one of the upstream primer and the downstream primer forms a primer. Identification of different pathotypes of plasmodiophora brassicae is rapidly completed through the PCR technology, and especially, the pathotype of plasmodiophora brassicae to cruciferae crops is rapidly and accurately identified through the difference of PCR amplification results; the identification result is accurate and repeatable, the operation process is simplified, the detection time is reduced, and the operation threshold is low.

This invention describes systems, primers, kits, and methods for detecting microsatellite instabilities in biological samples. Signal data is received from a capillary electrophoresis genetic analysis instrument, wherein the signal data is determined based on the fluorescence of fragments comprising nucleic acid sequences amplified from the biological sample by polymerase chain reaction (PCR). The nucleic acid sequences correspond to multiple different microsatellite loci and are obtained using multiple PCR primers configured to side-join the multiple microsatellite loci of the biological sample. When the PCR primers are combined with the biological sample and subjected to PCR amplification, fluorescently labeled DNA fragments comprising the multiple microsatellite loci are generated. The microsatellite instabilities of the biological sample are classified using the fluorescence data obtained from the multiple fluorescently labeled microsatellite loci.

The invention relates to the technical field of DNA library establishment and sequencing, and particularly discloses a method for performing full-coverage amplification on a DNA sample, a rapid DNA library establishment method, a sequencing method and application thereof. The amplification method comprises the following steps: carrying out low-temperature denaturation PCR (Polymerase Chain Reaction): contacting a DNA (Deoxyribonucleic Acid) sample with a plurality of groups of pre-amplification primer pairs, carrying out PCR amplification, and overlapping an amplification product of any one group of pre-amplification primer pairs in the plurality of groups of pre-amplification primer pairs and an amplification product of any adjacent group of pre-amplification primer pairs with at least 20 basic groups; the library building method comprises the amplification method. The sequencing method comprises the library building method. According to the sequencing method disclosed by the invention, the sequencing result can be quickly and efficiently obtained with high precision and high sequencing depth.

The invention discloses a digital PCR (Polymerase Chain Reaction) instrument. The excitation light unit and the bright field unit in the detection module are split into two mutually independent units, so that the excitation light unit and the bright field unit are respectively and independently arranged on the digital PCR instrument, and the installation difficulty of the detection module is reduced; according to the digital PCR instrument, the bright field unit and the exciting light unit are arranged in the detection module, and a connecting structure between the bright field unit and the exciting light unit is avoided, so that the structure of the detection module can be miniaturized, the overall structure of the digital PCR instrument is relatively compact and miniaturized, the exciting light unit and the bright field unit can be conveniently disassembled and assembled, and the detection efficiency is improved. Therefore, the bright field unit and the excitation light unit are repaired and maintained.

The utility model discloses an ultrasonic miniature stirring and mixing device which comprises a base, a sliding table is slidably mounted at the top of the base, a micro-fluidic chip is fixedly mounted above the sliding table, a plurality of PCR (polymerase chain reaction) tubes are mounted at the bottom of the micro-fluidic chip, a connecting seat is fixedly mounted at the top of the sliding table through an L-shaped support plate, and a plurality of PCR tubes are mounted at the bottom of the connecting seat. The connecting base is located between the micro-fluidic chip and the sliding table, the positions, close to the top ends, of the outer walls of the PCR tubes are each sleeved with an ultrasonic stirrer, and each ultrasonic stirrer comprises a shielding shell; according to the utility model, when the micro-fluidic chip is used for mixing experimental reagents in the PCR tube, the ultrasonic stirrer can be started, the reagents in the PCR tube can be mixed and stirred by utilizing the ultrasonic transducer, and the reagents in the PCR tube can also be mixed and stirred by utilizing the magnetic stirring mechanism, so that the stirring effect of the reagents is ensured, and the PCR tube does not need to be transferred; therefore, the experiment can be conveniently and quickly carried out, and the experiment efficiency is greatly improved.

The invention provides a qPCR (quantitative polymerase chain reaction) primer probe group for detecting various nodaviruses, a detection method and application, and belongs to the technical field of molecular biological detection. Aiming at various nodaviruses capable of infecting insect cell lines, including animal shed virus, Tn5 cell line virus, black beetle virus, borawa virus, nodavirus and macrobrachium rosenbergii nodavirus, a set of primer probe combination is designed. In the combination, part of viruses share the upstream primer and the probe, so that synchronous detection and identification of multiple target viruses in one reaction system are realized. The invention also provides a kit containing the primer probe group and a detection method. The detection method provided by the invention has the advantages of wide coverage range, high detection sensitivity, strong specificity, good matrix interference resistance and the like, and is suitable for safety detection and quality control of nodavirus pollution in insect cell banks, cell cultures and related biological products thereof.

The invention discloses a construction method of a glomerular IgA deposition mouse model, and relates to the technical field of animal model development and glomerular IgA dyeing. Comprising the following steps: obtaining Cas9 mRNA (messenger ribonucleic acid), guideRNA (ribonucleic acid) and donor DNA (deoxyribonucleic acid) in an in-vitro transcription manner; the method comprises the following steps: carrying out microinjection on Cas9 mRNA (messenger ribonucleic acid), gRNA (guide ribonucleic acid) and donor DNA (deoxyribonucleic acid) into fertilized eggs of a C57BL / 6J mouse; carrying out genotype identification on the F0 generation of birth mice through PCR (Polymerase Chain Reaction) amplification and sequencing; mating the F0-generation positive mouse with a wild type C57BL / 6J mouse, and breeding to obtain an F1-generation mouse; and spontaneous glomerular IgA deposition appears in 24 weeks after the F1 generation heterozygote mouse is born. According to the method, accurate mutation is introduced by utilizing a homologous recombination repair mechanism to construct a glomerular IgA deposition model, the efficiency is high, and the off-target risk is low.