29 results about "Gene expansion" patented technology

The present application relates to a kind of triplex-forming oligonucleotides (TFOs) sequence and its pharmaceutical application, especially to a kind of triplex-forming oligonucleotides sequence and its pharmaceutical application for inhibiting MDM2 and MDM4 gene amplification.The triplex-forming oligonucleotides sequence provided by the present application can form triplex structure locally in the amplified MDM2 / MDM4 gene of tumor cell, cause its replication stress, finally lead to the apoptosis of tumor cell.The triplex-forming oligonucleotides sequence provided by the present application and the medicine prepared using the sequence are aimed at the MDM2 / MDM4 amplification gene sequence of tumor rather than protein, have good drug resistance, and have no effect on normal cell without MDM2 / MDM4 gene amplification, have excellent safety, thus have good therapeutic effect on the tumor with MDM2 / MDM4 gene amplification.

The application discloses application of a peach fruit hexokinase PpHXK3 gene in preparation of a plant cold-resistant product, and is characterized in that the nucleotide sequence is shown as SEQ ID NO. 1, and the amino acid sequence is shown as SEQ ID NO. 2; a cloning method of PpHXK3 comprises the following steps: extracting total RNA of peach fruit and reversely transcribing the total RNA into cDNA as a template, and designing upper and lower amplification primers according to a gene sequence of PpHXK3; finally, a PpHXK3 gene amplification product is obtained through PCR amplification; the application of PpHXK3 in preparation of polygalacturonase PpPG21 inhibitors and / or polygalacturonase PpPG23 inhibitors, and the application of PpHXK3 in preparation of peach fruit cold-resistant products and heterologous plant cold-resistant products are also provided; and the advantages are that the decomposition of pectin is reduced, the integrity of a cell wall structure and the hardness of a fruit are maintained, the fruit has higher cold resistance, and cold damage in a storage and transportation process is reduced.

The invention relates to the technical field of gene detection, in particular to a rapid shrimp iridovirus detection primer group based on an RPA-CRISPR / Cas12a system, application and a kit. The primer group comprises an iridovirus constant-temperature gene amplification primer and a specific recognition amplification product target sequence probe; wherein the iridovirus constant-temperature gene amplification primer, the specific recognition amplification product target sequence CRISPR probe and the ssDNA reporter probe are recorded in the specification. An isothermal amplification primer and CrRNA are designed based on the shrimp red color virus gene, the target gene is amplified through the isothermal amplification technology, Cas12a is guided by the CrRNA to accurately recognize and activate trans-cleavage, a test strip report probe is synchronously split, and the result is interpreted by naked eyes within 30 min by virtue of the visual advantage of an immune test strip. The whole process does not need large instruments, the operation is simplified, the cost is low, and a visual and efficient solution is provided for on-site rapid diagnosis.

The invention discloses an SNP (Single Nucleotide Polymorphism) site, a DNA (Deoxyribose Nucleic Acid) fragment, a molecular marker primer and a method for identifying the color of cerasus humilis flesh, the SNP site is located at the 422nd basic group site of a cerasus humilis DFR gene, the polymorphism is T / G, the SNP site of red flesh is T, and the SNP site of yellow flesh is G. The nucleotide sequence of the DNA fragment is as shown in SEQ ID NO.1 or SEQ ID NO.2, and the molecular marker primer comprises a forward primer as shown in SEQ ID NO.3 and a reverse primer as shown in SEQ ID NO.4. The molecular marker primer has the advantages that the nucleotide sequence of the DNA fragment is as shown in SEQ ID NO.1 or SEQ ID NO.2; the method comprises the following steps: (1) collecting a to-be-detected cerasus humilis plant tissue sample and extracting total RNA (Ribonucleic Acid); (2) carrying out reverse transcription on the total RNA to obtain cDNA (complementary deoxyribonucleic acid); step (3), amplifying the cerasus humilis DFR gene by using the molecular marker primer by taking the cDNA as a template; (4) purifying and sequencing an amplification product; and (5) judging whether the pulp of the to-be-detected cerasus humilis sample is red or yellow according to a sequencing result. The technical problems that a traditional cerasus humilis pulp color identification method is long in period and not beneficial to popularization can be solved.

1. The name of the design product: Gradient gene amplification instrument (gradient PCR instrument). 2. The use of the design product: for gene amplification reaction. 3. The design points of the design product: in the combination of shape and pattern. 4. The picture or photo that best indicates the design points: perspective view.

This automated gene testing device enables accurate and rapid determination of gene amplification reactions within a reaction vessel through precise dispensing operations controlled by the rotation of a microfluidic chip and precise heating operations controlled by the temperature of the microfluidic chip. It allows for simple and rapid genetic testing of multiple viruses and other pathogens at the genetic level, even in on-site settings, without requiring special expertise or skills. [Solution] A gene amplification reaction is performed using a disc-shaped test chip equipped with a microfluidic centrifugal dispensing function, comprising a liquid delivery unit, a heating unit, an imaging unit, a display unit, and a control unit. The test chip comprises an injection reservoir set with a liquid sample and multiple reaction chambers set with primers. The liquid delivery unit comprises a drive source and a rotating shaft, and the heating unit comprises a heater and a lifting device. The imaging unit comprises a light source and a camera, and the display unit comprises a determination device and a display device.

The invention provides a molecular diagnosis method for avian leukosis virus and Marek's disease virus mixed infection. The molecular diagnosis method comprises the following steps: (S01) collecting diseased material tissues; (S02) carrying out nucleic acid extraction on the sick material tissue of the sick chicken; (S03) detecting the nucleic acid by using specific primers of avian leukosis and Marek's disease; (S04) preparing a PCR (Polymerase Chain Reaction) detection system, and respectively adding ALV-env gene amplification primers and MDV-meq gene amplification primers for gene amplification; (S05) carrying out 1% agarose gel electrophoresis detection on the amplification product in the step (4), and carrying out sequence determination after determining a target band; (S06) carrying out BLAST comparison on the sequences obtained by sequencing, carrying out gene sequence analysis and drawing an evolutionary tree; and (S07) determining the virus type according to the homology and the evolutionary branches, so as to realize rapid diagnosis of ALV and MDV mixed infection, strain characteristic analysis and exclusion of other similar epidemic diseases, and provide technical support for accurate prevention and control of AL and MD in the poultry industry, provenance purification and epidemiological investigation.

The application discloses a gene amplification detection method based on nucleic acid mass spectrometry technology, comprising the following steps: setting a target gene region, configuring a reference gene, and confirming experimental primers based on the target gene region and the reference gene; obtaining an experimental sample, performing a primer reaction processing operation based on the experimental primers and the experimental sample, and obtaining a sample to be analyzed; setting an interpretation interval, configuring a sample applicator and a nucleic acid mass spectrometer, performing a data analysis and interpretation operation based on the sample to be analyzed, the interpretation interval, the sample applicator and the nucleic acid mass spectrometer, and obtaining a gene amplification interpretation result; the application can achieve a detection accuracy higher than that of the fluorescence in situ hybridization technology and the immunohistochemical technology in the prior art, a detection throughput higher than that of the fluorescent quantitative PCR technology and the digital PCR technology, a detection cost lower than that of the fluorescent quantitative PCR technology and the digital PCR technology, and a detection time shorter than that of the second-generation sequencing NGS technology.

The application discloses a kind of binarization observation methods and systems of gene network expression state, it is related to the field of life science cell gene expression determination technical field, method includes: according to gene dictionary and the gene dimension of measured cell determines random measurement matrix, and then determines the multiple gene name groups corresponding to random measurement matrix;For each gene name group corresponding to the random measurement matrix, T7 is used to customize positive value primer and negative value primer respectively, and is labeled respectively, to obtain corresponding positive value labeled primer and negative value labeled primer;Using positive value labeled primer, the gene data corresponding to positive value gene name group is amplified, and the gene data corresponding to negative value gene name group is amplified using negative value labeled primer, to obtain comprehensive gene amplification data, and it is labeled to obtain gene observation data;According to gene observation data and gene dictionary, the gene expression of measured cell is calculated.The application improves gene expression measurement stability.

The invention relates to the technical field of gene amplification, in particular to a PCR gene amplification device which comprises a shell, an amplification groove is formed in the upper portion of the shell, a heat conduction plate is fixedly installed at the bottom of the amplification groove and penetrates to the lower portion of the amplification groove, a semiconductor Peltier is installed below the heat conduction plate in an attached mode, and a water cooling structure is arranged below the semiconductor Peltier. The water cooling structure comprises a water cooling box fixedly installed below the semiconductor Peltier and a heat dissipation piece fixedly installed on one side of the shell, a water inlet pipe and a water outlet pipe are arranged on the water cooling box, a storage piece is connected to the water outlet pipe in parallel, and the storage piece is used for conducting heat preservation storage on liquid flowing out of the water outlet pipe. According to the semiconductor Peltier element, a cold source or a heat source generated below the semiconductor Peltier element can be fully utilized, and the cold source or the heat source is not directly discharged.

The invention discloses a primer group, a kit and a method for identifying deer species based on IGF1 (Insulin-like Growth Factor 1), IGF2 (Insulin-like Growth Factor 2) and RAF1 genes. The primer group comprises three pairs of specific primers aiming at IGF1, IGF2 and RAF1 genes, and the sequences of the three pairs of specific primers are respectively SEQ ID NO: 12 and 13, SEQ ID NO: 14 and 15, and SEQ ID NO: 16 and 17. In specific identification, DNA of a sample is extracted, PCR amplification is carried out by using the primer group, and then the fragment length difference of an amplification product is analyzed through agarose gel electrophoresis. By comparing specific length combinations of IGF1, IGF2 and RAF1 gene amplified fragments, the four types of cervidae animals can be accurately distinguished. According to the application, the identification system for the cervidae species based on the IGF1, IGF2 and RAF1 genes is established for the first time, and the identification system has the advantages of rapidness, accuracy, low cost and no need of sequencing, and is suitable for species source identification and market supervision of related products.

A method for predicting responsiveness to a HER2-directed therapy by assessing HER2 heterogeneity in a tumor includes contacting a sample of the tumor with a biomarker-specific reagent that specifically binds to HER2 protein and detecting HER2 protein in the sample, contacting the sample of the tumor with a first nucleic acid probe that specifically binds HER2 genomic DNA and detecting HER2 gene amplification status in the sample, contacting the sample of the tumor with a second nucleic acid probe that specifically binds HER2 RNA and detecting HER2 RNA status in the sample scoring the HER2 protein (IHC), HER2 gene (DISH), and HER2 RNA (RNA-ISH), predicting that the tumor is responsive to the HER2-directed therapy if the tumor reveals a first foci having a first score and a second score, in which the first score and the second score are not the same.

The invention discloses a detection method for noninvasive prenatal detection of fetal trisomy 21 syndrome based on digital PCR, and relates to the technical field of digital PCR. According to the invention, a target area detection amplification primer and a Taqman-MGB probe are designed according to different sequences of human chromosome 21 genes, and a specific common amplification primer and a specific Taqman-MGB probe are designed according to similar sequences of the human chromosome 21 genes. Accurate detection of a low-proportion trisomy 21 syndrome positive sample is realized, and the omission ratio is greatly reduced. Compared with next-generation sequencing, multiple qPCR technologies and the like, the detection method provided by the invention has the advantages of simplicity in operation, simplicity in data analysis, short detection period, low cost and the like, meanwhile, by innovatively using a similar sequence design technology, high consistency of amplification efficiency of a detection gene and a reference gene is ensured, and the detection accuracy is improved. The detection sensitivity and accuracy are obviously improved.

The invention provides a detection method for detecting Sf9 host DNA residues by using a fluorescent probe and application, and belongs to the technical field of biological detection.According to the method, a specific primer pair (such as SEQIDNO: 1 / 2) and the fluorescent probe (such as SEQIDNO: 11) are designed according to a high-copy conserved sequence of Sf9 host DNA, a kit containing a gene amplification solution, a standard substance and a quality control substance is matched, DNA is extracted through a liquid nitrogen grinding-column extraction method, and the detection result is accurate. And carrying out real-time fluorescent quantitative PCR amplification and then carrying out detection. According to the method, the detection sensitivity reaches 0.003 pg / mu L, the linear range is 300 pg / mu L-0. 003 pg / mu L, cross reaction to irrelevant DNA is avoided, the repeatability CV% is smaller than or equal to 10%, the reagent is still stable after being frozen and thawed for 10 times, and the whole process is completed within 1.5 hours; the method solves the problems of insufficient specificity and sensitivity, tedious operation and the like of the existing method, and can be used for quality control of biological products such as gene therapy drugs and cell vaccines.

The purpose of the present invention is to establish a simple, efficient, and rapid genetic testing method for bacteria in clinical settings, and to provide: a gene amplification method for specifically detecting and evaluating the amount of bacteria associated with periodontal disease and caries; a primer set for a gene amplification reaction; a kit for detecting genes of bacteria that cause periodontal disease or caries, the kit including the primer set; and a kit for evaluating the severity of periodontal disease or caries. The present invention is a method for detecting oral bacteria that are present in plaque or saliva above or below the gingival margin, or bacteria that cause periodontal disease or caries, by detecting genes of the oral bacteria or causative bacteria using a gene amplification method, the detection method being characterized in that (1) annealing and extension reactions in the gene amplification method are implemented at substantially the same temperature, and / or (2) the gene length of a target gene as an amplification product is short.

This application relates to a molecular diagnostic reference for human gene detection, its preparation method, and its application. The reference is a recombinant cell, and the chromosome of the recombinant cell integrates the exogenous human variant gene to be detected in the form of homologous recombination; wherein the recombinant cell is selected from either recombinant mammalian cells or recombinant insect cells. This reference is substantially similar to clinical samples or cell lines of the human variant gene to be detected in terms of structure, performance, and composition, and can fully simulate gene deletion, gene insertion, gene fusion, gene amplification, fragment duplication, gene breakage, SNP, and gene overexpression. Furthermore, the preparation method of the reference is simple to operate, has a short preparation cycle, produces large quantities, has low production costs, and its source and performance are sufficiently stable. During use, the entire process from sample processing to detection can be monitored, exhibiting high biosafety and making it well-suited for application in molecular diagnostic products for human gene detection.

The invention belongs to the technical field of bioengineering, and particularly relates to a recombinant plasmid for promoting synthesis of tylosin, an engineering bacterium and a preparation method thereof, the genetically engineered bacterium (Streptomyces fradiae G3-F2) is preserved in China Center for Type Culture Collection on February 24, 2025, and the preservation number is CCTCC NO: M 2025278. According to the streptomyces fradiae G3-F2 disclosed by the invention, through site-specific recombination, a recombinant plasmid is constructed by utilizing an optimized ZouA gene DNA amplification system, so that multi-copy increase of a tylF gene is realized, and further, streptomyces fradiae G3-F2 is prepared; the enzymatic reaction rate of the methyltransferase is greatly improved; the efficient conversion of the component C is realized; and low-temperature culture fermentation is combined, so that the tylosin synthesis rate and efficiency are greatly improved. The titer of the strain in a 50L tank can reach 18500U / mL under the low-temperature culture condition of 28 DEG C, so that the production cost can be greatly reduced.

The utility model relates to a gene amplification diagnostic kit which comprises a bottom plate, two limiting plates are fixedly connected to the position, close to the left side, of the top of the bottom plate, the two limiting plates are symmetrically arranged front and back, a mounting plate is fixedly connected to the position, close to the bottom, of the left side of the bottom plate, and an air cylinder is mounted in an inner cavity of the mounting plate; a power output shaft of the air cylinder extends to an inner cavity of the bottom plate, and two movable plates are attached to the bottom of the bottom plate. Through mutual cooperation of a bottom plate, an air cylinder, a mounting plate, a limiting plate, a constant-temperature box, a movable plate, an overturning plate, a fixed plate, a movable plate, an L-shaped fixed rod, a placement plate, a top plate, a connecting plate, a kit, a test tube, a transverse plate, a first rotating shaft, a middle plate, a second rotating shaft, a sliding block, a hinge block, a crank and other components, the constant-temperature box and the kit can be combined together; the kit can quickly enter the inner cavity of the constant-temperature box, so that the test efficiency is improved.

This invention belongs to the field of bioengineering technology, specifically relating to a recombinant plasmid for tylosin synthesis, an engineered bacterial strain, and its preparation method. The genetically engineered bacterial strain (Streptomyces freundii G3-F2) was deposited at the China Center for Type Culture Collection (CCTCC) on February 24, 2025, with accession number CCTCC NO: M 2025278. This invention achieves site-specific recombination, utilizes an optimized ZouA gene DNA amplification system to construct the recombinant plasmid, and realizes… tulle Multiple copies of the F gene were increased, resulting in the preparation of *Streptomyces freundii* G3-F2. This significantly improved the rate of methyltransferase enzymatic reaction and achieved efficient conversion of component C. Combined with low-temperature fermentation, the rate and efficiency of tylosin synthesis were greatly enhanced. The strain achieved a titer of 18500 U / mL in a 50L tank under 28℃ low-temperature culture conditions, which can significantly reduce production costs.

The present invention relates to an RPA primer set for identifying Hanwoo, a composition including the primer set, a kit including the composition, and a method for identifying Hanwoo using the primer set. According to one aspect of the present invention, the primer set, the composition including the primer set, the kit including the composition, and the method for identifying Hanwoo using the primer set can rapidly amplify specific nucleotide sequences using a single-strand DNA binding (SSB) and a recombinase as an RPA method. Since the reaction is possible under isothermal conditions, it is possible to specifically distinguish between Hanwoo and imported cattle (non-Hanwoo) simply by the presence or absence of a gene amplification product without the need for temperature-circulating equipment such as a PCR machine. Furthermore, it has excellent effects that allow for rapid and accurate on-site identification with only a small amount of sample, for example, about 25 mg, which can be utilized for origin labeling management.

The invention belongs to the technical field of microbiological detection, and particularly relates to a primer and method for detecting streptococcus suis and application. The nucleotide sequences of the primer for detecting the streptococcus suis are shown as SEQ ID NO.1 and SEQ ID NO.2, and the target gene is recA; the nucleotide sequences of the primers for detecting the virulence gene of the streptococcus suis are shown as SEQ ID NO.3 and SEQ ID NO.4, and the virulence gene is sly. The detection gene amplification primers do not influence each other, so that the accuracy and reliability of the detection gene amplification primers can be ensured.

The invention relates to the technical field of gene detection and risk assessment, and particularly discloses a portable single gene expression rapid detection system and a tumor risk assessment method thereof. Calling a temperature control time sequence parameter and an amplification efficiency reference threshold value matched with the target gene from a preset database; the reaction cavity is controlled to be in different temperature intervals in sequence according to the temperature control time sequence parameters so as to execute an amplification reaction, in the reaction process, fluorescence intensity signals in the reaction cavity are collected according to the preset sampling frequency, and the fluorescence intensity change rate is calculated based on the time dimension. A closed-loop intelligent detection and evaluation system is constructed by integrating a sample physical property pre-detection module, an amplification efficiency real-time prediction module, a dynamic compensation module and the like; the system can realize multi-dimensional active monitoring and interference suppression of the whole process of gene amplification reaction on a portable platform.

A gene amplification chip may include: a cover layer having a solution inlet through which a sample solution to be injected; a chamber layer disposed on one surface of the cover layer, and having a chamber to receive the sample solution when the sample solution is injected through the solution inlet such that an amplification reaction of the sample solution occurs in the chamber; a bottom layer disposed on another surface of the chamber layer; and a photothermal film attached to an outer surface of the bottom layer, and configured to convert light into heat to heat the sample solution received in the chamber.

This invention belongs to the field of microbial detection technology and provides a high-throughput, low-cost targeted next-generation sequencing method, including sample collection and preprocessing, target gene amplification, direct library construction, multi-sample pooling, high-throughput sequencing, and data analysis. By optimizing the sample collection and preprocessing, target gene-specific amplification, direct library construction, multi-sample pooling, and high-throughput sequencing and data analysis processes, this invention significantly reduces sequencing costs and improves detection efficiency, while ensuring data accuracy and reliability. It effectively solves the problems of high cost, complex operation, large data volume requirements, and unstable results in existing technologies, and is particularly suitable for low-cost, multi-sample simultaneous detection needs in the field of microbial detection.

The invention relates to the technical field of molecular biology detection, and discloses a primer probe combination for verifying whether gene mutation occurs in a true gene or not and application of the primer probe combination, and the primer probe combination comprises the following components: a forward primer, a reverse primer and a Blocker. Sequence overlapping exists between the Blocker and the forward primer, and the Blocker is designed to be complementary with the homologous gene and not complementary with the true gene at the difference site of the true gene and the homologous gene, so that the Blocker and the forward primer are competitively combined with the genome DNA, and amplification of the homologous gene is inhibited. And performing Sanger sequencing on the amplification product, comparing mutation sites and difference sites of the true gene and the homologous gene in a sequencing result, and judging whether the gene mutation occurs on the true gene or not. The established method successfully verifies that the prenatal fetal sample SLC6A8 NM005629.4 c.1540 Cgt is used for detecting the prenatal fetal sample SLC6A8 NM005629.4 T (p.R < 514 * >) and SDHA NM004168.4 c.433 Cgt are added into the kit; t (p.Q145 *) mutation authenticity can be realized, and variation sources can be inferred based on family analysis. The method has the advantages of simplicity and convenience in operation, high specificity, low cost, facilitation of popularization and the like.

A method for predicting responsiveness to a HER2-directed therapy by assessing HER2 heterogeneity in a tumor includes contacting a sample of the tumor with a biomarker-specific reagent that specifically binds to HER2 protein and detecting HER2 protein in the sample, contacting the sample of the tumor with a first nucleic acid probe that specifically binds HER2 genomic DNA and detecting HER2 gene amplification status in the sample, contacting the sample of the tumor with a second nucleic acid probe that specifically binds HER2 RNA and detecting HER2 RNA status in the sample scoring the HER2 protein (IHC), HER2 gene (DISH), and HER2 RNA (RNA-ISH), predicting that the tumor is responsive to the HER2-directed therapy if the tumor reveals a first foci having a first score and a second score, in which the first score and the second score are not the same.

The invention relates to a constant-pressure heat cover device for a gene amplification instrument, and the device comprises a heat cover assembly which comprises at least one installation cavity used for placing a reaction tube; the heat cover bracket comprises a first connecting part and a second connecting part, and the first connecting part is movably connected with the heat cover assembly; the rotating piece is rotatably arranged above the mounting cavity and is connected between the two first connecting parts; the moving part is arranged on one side of the rotating part and is movably connected with the second connecting part; the moving part is provided with a first inclined surface, and the first inclined surface is used for abutting against the rotating part; the first elastic piece is arranged between the hot cover bracket and the rotating piece; when the heat cover assembly is pressed on the reaction tube, the reaction tube reversely pushes the rotating part to rotate and move in the first direction, and pushes the moving part to move in the second direction. The floating rotating part is in contact with the reaction tube to drive the moving part with the inclined surface to move; the first elastic piece is compressed to generate constant pressure, dimensional changes caused by tolerance, thermal expansion and the like are absorbed, and the pressure is kept constant.

The present invention provides a PCR plate, a nucleic acid extraction cartridge, and a polymerization enzyme chain reaction apparatus containing the same, which can perform nucleic acid extraction and real-time gene amplification testing fully automatically, can test multiple targets in a single operation, are easy to use, and in particular can provide accurate results in a short time. [Solution] The PCR plate includes a body portion having one or more reaction wells; an insertion portion extending from the body portion and having an injection port into which nucleic acid solution is injected, which is inserted into a nucleic acid extraction cartridge; a flow channel portion that allows the nucleic acid solution to flow from the injection port into the reaction wells; and a blocking portion attached to the body portion that blocks the backflow of nucleic acid solution from the reaction wells to the flow channel portion.

The invention provides a FISH probe combination and kit for detecting NECTIN4 gene amplification and a preparation method and application of the FISH probe combination and kit. The FISH probe combination comprises an NECTIN4 gene probe and a first chromosome internal reference probe; a preparation template of the NECTIN4 gene probe is composed of any one of the following BAC clone plasmids (1) to (3): (1) the BAC clone plasmids are RP11-157H6, CTD-3003O12 and RP11-4N6, and (2) the BAC clone plasmids are RP11-157H6, CTD-3003O12 and RP11-4N6; (2) cloning plasmids CTD-2299J15 and CTD-2502B20 by using a BAC (Bacterial Amplification Cellulose); and (3) cloning the plasmid CTD-2502B20 by using the BAC. The probe combination can detect the amplification condition of the NECTIN4 gene of a patient, provides data support for clinical tests and individualized treatment, has the characteristics of high sensitivity and good specificity, and can rapidly hybridize within 1 hour and stably interpret the signal intensity at high energy.