13 results about "Gene specific primer" patented technology

The invention discloses a megalurothrips megalurothrips methylamino abamectin benzoate resistance molecule detection method which comprises the following steps: (1) collecting megalurothrips megalurothrips female adults in a plurality of regions, and extracting DNA (deoxyribonucleic acid) in a single region in a mixed manner; (2) designing glutamic acid gated chloride ion channel gene specific primers of the megalurothrips megalurothrips, respectively adding specific tags at the front ends of the primers in all regions, and carrying out PCR (Polymerase Chain Reaction) amplification; mixing the PCR products of each region to obtain a library; (3) performing second-generation amplicon sequencing on the library to obtain off-machine data; and (4) analyzing offline data, merging the data in the same region, calculating the mutation frequency of each region by taking the ratio of the mutation (G232S) of the target sequence as an index, and judging the resistance of the megalurothrips usitatus to the emamectin benzoate. The method can be used for rapidly and massively monitoring the emamectin benzoate resistance condition of the field megalurothrips megalurothrips, and an important basis is provided for high-level emamectin benzoate resistance early warning and megalurothrips megalurothrips chemical treatment.

The invention relates to the technical field of molecular biology, in particular to a sheep polyfetal gene FecB specific LAMP primer pair and a sheep polyfetal gene FecB genotype detection method. According to the invention, the visual detection method of the sheep polyfetal gene FecB genotype is established based on the LAMP technology, large expensive equipment is not needed in the detection process, and the detection cost is remarkably reduced; the detection time is shortened to about 20 minutes, so that the detection efficiency is greatly improved; in addition, the detection result is visual, intuitive and readable, and the method is particularly suitable for field detection, shows great practical value and popularization potential, and has wide application prospects. The invention provides a new technical means for efficiently constructing a sheep breeding dominant genetic resource group, and is beneficial to accelerating a marker-assisted selection breeding process of sheep breeding traits.

The present invention relates to a method for detecting the rcAAV content in a sample of recombinant adeno-associated viral vector, said method being characterized by the fact that the cells are cultivated using cell culture flasks with a capacity of 4 to 5 mL and are infected three times or more without loss of rcAAV, then the rcAAV content in the sample of recombinant adeno-associated viral vector is obtained by performing a quantitative PCR on the ultrafiltration concentrates of the first and last infection harvests using the rep2 gene-specific primers shown in SEQ ID NO: 1 and 2 and the Taqman probe shown in SEQ ID NO: 3. The present invention also relates to a kit for performing the method for detecting the rcAAV content in a sample of recombinant adeno-associated viral vector according to the invention.

The invention belongs to the technical field of tumor molecule detection, and particularly discloses a method and a kit for detecting HER2 gene mRNA (messenger Ribonucleic Acid) trace expression based on an RT-ddPCR (Reverse Transcription-Derived Polymerase Chain Reaction) technology. The kit contains a cross HER2 exon 19-20 / 20-21 specific primer probe, a reference gene specific primer probe and reverse transcriptase, and the ddPCR premix solution, the HER2 probe and the reference gene probe are respectively marked with different fluorophores. According to the detection method, after RNA extraction, reverse transcription and RT-ddPCR amplification of a detection sample are carried out, the HER2 / internal reference copy number ratio is calculated to realize typing, a breast cancer sample is completely negative when the Ratio is smaller than 1.0, is ultralow in expression when the Ratio is larger than or equal to 1.0 and smaller than 5.0, is low in expression when the Ratio is larger than or equal to 5.0 and smaller than 50.0, and is medium-high in expression when the Ratio is larger than or equal to 50.0. The detection sensitivity reaches 0.1 copy / microliter, absolute quantification can be achieved, adaptability to FFPE samples is high, breast cancer patients with ultralow expression of HER2 in IHC 0 can be accurately screened, a molecular basis is provided for accurate medication of ADC drugs such as Trastuzumab deruxtecan, and the kit can also be used for related detection of solid tumors such as gastric cancer and cervical cancer with HER2 gene amplification as a marker.

The invention discloses a digital PCR detection system, kit and method for detecting single nucleotide polymorphism and copy number variation of a CYP2D6 gene and application, and belongs to the technical field of molecular biology. The method comprises the following steps: designing three pairs of CYP2D6 and reference gene RPP30 specific primers, matching with four locked nucleic acid modified TaqMan fluorescent probes, constructing a single-tube multiple digital PCR detection system, and synchronously and accurately detecting three single nucleotide polymorphisms of CYP2D6 * 10, CYP2D6 * 36 and CYP2D6 * 41 and CYP2D6 * 5 copy number variation; wherein the RPP30 reference gene can synchronously realize sample amplification quality control and target gene absolute quantitative calibration. The detection system depends on the advantages of high specificity of the LNA probe and high sensitivity of digital PCR, the detection sensitivity reaches 0.02 ng / mu L, the detection period is short, and the result is stable and reliable.

The invention relates to a fluorescent quantitative PCR kit and method for detecting a mycobacterium tuberculosis complex group in a tongue swab, and belongs to the field of pathogenic microorganism detection, the fluorescent quantitative PCR kit comprises the following components: an amplification reaction solution, a primer probe mixed solution and an external reference; the amplification reaction solution is a PCR (Polymerase Chain Reaction) buffer solution containing Taq DNA (Deoxyribose Nucleic Acid) polymerase, Mg < 2 + > ions, dNTP (dUTP) and UDG enzyme; the primer probe mixed solution comprises an IS6110 specific inner primer pair, an outer primer pair and a probe; the kit comprises an IS1081 specific inner primer pair, an IS1081 specific outer primer pair and a probe, a bacillus subtilis specific primer pair and a probe; the invention discloses a human gene specific primer pair and a probe. The fluorescent quantitative PCR kit disclosed by the invention can be used for rapidly and sensitively detecting the mycobacterium tuberculosis complex DNA in a tongue swab sample of a tuberculosis patient, has the advantages of high sensitivity and high specificity, and has a wide application prospect.

The application discloses a W-specific gene for identifying female and male of each instar of Ostrinia furnacalis, a specific primer pair, a kit and a detection method. The nucleotide sequence of the W-specific gene is shown as SEQ ID NO:1. The application also provides a specific primer pair for detecting the W-specific gene. The application further provides a PCR detection method for rapidly identifying female and male of each instar of Ostrinia furnacalis. After gDNA of Ostrinia furnacalis is extracted, PCR amplification is carried out, and agarose gel electrophoresis is carried out on the PCR amplification product. The cloned fragment of the W-specific gene of the female has a corresponding band in the agarose gel electrophoresis diagram, and the male has no corresponding band. The method is suitable for identifying female and male of each growth and development stage of Ostrinia furnacalis, has short detection time, simple operation and low cost, and is suitable for large-scale detection. The method has high accuracy, and non-insect taxonomists can also identify female and male of each growth and development stage of Ostrinia furnacalis through the method.

The invention provides an amplification library building method for high-throughput sequencing verification of synthetic genes. The amplification library building method comprises the following steps: obtaining a joint connection product of which two ends are connected with double-chain joints after a to-be-detected synthetic gene sequence, and carrying out first PCR (Polymerase Chain Reaction) amplification by adopting a first gene specific primer by taking the joint connection product as a template to obtain a first amplification product; taking the first amplification product as a template, and carrying out second PCR amplification by adopting a second gene specific primer to obtain a second amplification product; performing thermal denaturation single-chain separation and single-chain cyclization on the second amplification product to obtain single-chain circular DNA; the preparation method comprises the following steps: carrying out DNB preparation on single-stranded circular DNA to obtain DNB nanospheres; and sequencing the DNB nanospheres to obtain synthetic gene sequence information. The method verifies that the synthesized gene is high in flux, and the flux requirement of large-scale gene synthesis can be met.

The present application relates to the technical field of virus detection, in particular to a primer probe combination for feline infectious peritonitis virus serotyping, application and kit thereof. The primer probe combination comprises FIPV N gene specific primer probe, FIPV I serotype S gene specific primer probe and FIPV II serotype S gene specific primer probe. The three groups of primer probes can be used for serotyping identification of FIPV I serotype strain and FIPV II serotype strain. The primer probe combination can be used for differential diagnosis of different serotypes of FIPV by fluorescence quantitative PCR method, and has the advantages of accurate identification, high sensitivity, simple operation, short time consumption and the like, and can provide effective technical means for rapid diagnosis of feline infectious peritonitis virus in China and prevention of feline infectious peritonitis.

The invention relates to the technical field of molecular biology, in particular to a detection method of a sheep multi-fetal gene FecB genotype and an identification method of sheep multi-fetal traits. The specific LAMP primer pair of the sheep multi-fetal gene FecB is designed, the detection method of the genotype of the sheep multi-fetal gene FecB is established based on the primer pair, large-scale detection of single nucleotide polymorphic sites of the FecB gene can be realized, and the sheep multi-fetal traits can be identified according to the detection result. The method has the advantages of simplicity in operation, high efficiency, rapidness and high sensitivity, provides an efficient and convenient detection method for early selection and molecular breeding of sheep, and has a wide application prospect.

The invention relates to the technical field of molecular biology, in particular to a method for detecting the genotype of a sheep multi-fetal major gene FecB based on an isothermal amplification technology. The invention provides the specific RPA primer group for the sheep multi-fetal major gene FecB, and a novel method for typing the polymorphic sites of the sheep multi-fetal major gene FecB is established by utilizing an RPA technology, and the method has the advantages of simplicity in operation, high efficiency, rapidness and high accuracy; genetic typing of the sheep multi-fetal major gene FecB and identification of sheep multi-fetal character individuals can be rapidly achieved on a large scale, a pasture is helped to conduct early selection on high-reproduction breeding stock, the group reproduction efficiency can be effectively improved, genetic improvement of the lambing number, the lambing rate and other characters of a sheep flock can be accelerated, continuous optimization of the population is achieved, and the method has wide application prospects.

The application provides a kind of mitochondrial feature sequence, specific primer, kit and method for identifying Chinese saprobic boletus, belongs to boletus identification technical field, the mitochondrial feature sequence is as shown in SEQ ID NO.1, the mitochondrial gene specific primer includes upstream primer and downstream primer;The nucleotide sequence of the upstream primer is as shown in SEQ ID NO.2;The nucleotide sequence of the downstream primer is as shown in SEQ ID NO.3.The kit includes the mitochondrial gene specific primer and reference primer of the application.By designing specific primer pair on the specific region of Chinese saprobic boletus mitochondrial gene, combined with reference primer, the rapid and accurate identification of Chinese saprobic boletus is realized.

This invention discloses a dual-gene specific primer set for rapid identification of Chiroptera species and its application method. The primer set includes primer pair COI-F / COI-R for amplifying the mitochondrial COI gene and primer pair Cytb-F / Cytb-R for amplifying the Cytb gene, with nucleotide sequences shown in SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, and SEQ ID NO.4, respectively. Experiments have demonstrated that this primer set exhibits good interspecific specificity and intraspecific conservation, effectively distinguishing Chiroptera from other closely related mammals and accurately identifying cryptic species and juvenile samples that are difficult to distinguish morphologically. This invention employs dual-gene markers for mutual verification, significantly improving the accuracy and reliability of identification, providing a powerful molecular biology tool for rapid and accurate identification of Chiroptera species, and has significant application value in fields such as biodiversity surveys, disease tracing, and ecological conservation.