837 results about "Specific primers" patented technology

The invention discloses SSR (simple sequence repeat) multiple PCR (polymerase chain reaction) primers for paternity test of grass carp and application of the SSR multiple PCR primers. The primers comprise 16 pairs of specific primers, and base sequences of the 16 pairs of specific primers are respectively shown as SEQ ID NO.1-32 in sequence. The invention further discloses a fish paternity test kit comprising the SSR multiple PCR primer, an SSR multiple fluorescent PCR method for grass carp paternity test and application of the SSR multiple PCR primer, the kit or the method in grass carp paternity test. According to the kit, 16 pairs of specific SSR primers, multiple fluorescent PCR and a universal amplification primer technology are combined, 16 sites can be detected at a time, universal primers can be repeatedly used, and compared with simple single-site detection, the efficiency is improved, and the cost is reduced; the microsatellite loci contained in the invention are 3-6 basic groups, the allele size interpretation is more accurate, and the accuracy of genotype data is improved.

The invention discloses protease for improving the resistance of potatoes to late blight as well as a coding gene and application thereof, and belongs to the technical field of genetic engineering and plant disease resistance breeding. The amino acid sequence of the protease StRD21 is as shown in SEQ ID NO.2, or the protease StRD21 has a derivative sequence with the same disease-resistant function. The nucleotide sequence of the coding gene is as shown in SEQ ID NO. 1. The invention also provides a specific primer pair for cloning the gene, a potato recombinant overexpression vector containing the gene and a transgenic host cell. The StRD21 gene is introduced into potatoes and overexpressed, so that the resistance of the potatoes to late blight can be remarkably enhanced. The invention provides an effective gene resource and a biotechnological means for cultivating a new variety of disease-resistant potatoes.

The invention discloses a nucleic acid product, a kit and a method for detecting various pathogens. The nucleic acid product comprises a primer pair and a probe, wherein the nucleotide sequences of the primer pair and the probe are respectively shown as SEQ ID NO: 1-24. Specific primer probes are respectively designed for conserved regions of eight respiratory tract pathogenic bacteria, synchronous detection of multiple pathogens is realized through a one-tube eight-color fluorescent PCR, freeze-drying and ultrasonic direct diffusion method technology, experimental operation steps are remarkably reduced, the detection time is shortened, the result interpretation complexity is reduced, and in addition, the detection efficiency is greatly improved. The anti-interference capability is strong, and the sensitivity is high.

The invention relates to the technical field of microbiological detection and molecular biology, in particular to a specific detection primer for bifidobacterium longum subsp. Longum BBMN68 and application of the specific detection primer. The specific primer for detecting the bifidobacterium longum subsp.longum BBMN68, provided by the invention, comprises an upstream primer and a downstream primer, and nucleotide sequences are shown as SEQ ID NO.7-8. The invention also provides a kit for detecting the bifidobacterium longum subsp.longum BBMN68. The specific DNA fragment provided by the invention is a specific molecular marker of BBMN68, and the specific primer is designed aiming at the fragment sequence, so that the specific amplification of the strain can be realized, no cross reaction is caused to other related strains, the detection sensitivity is higher, and the detection requirements of samples with different concentrations can be met; the method can be used for specific detection and accurate quantification of the BBMN68 strain in yoghourt and other products, and has good practical application value.

The invention relates to the technical field of biological detection, and discloses a mycoplasma pneumoniae nucleic acid rapid detection method, which comprises the following steps: mixing a sample to be detected with a multifunctional pretreatment buffer solution containing tris (hydroxymethyl) aminomethane, potassium acetate, polyethylene glycol, saponin, spermine tetrahydrochloride and tris (2-carboxyethyl) phosphine hydrochloride, and carrying out cracking treatment; a cracking mixed solution is obtained; redissolving the enzymatic isothermal amplification basic reagent, the specific primer and the probe to obtain a redissolving amplification system; sucking the supernatant of the cracking mixed solution, adding the supernatant into a redissolving amplification system, and adding a magnesium acetate solution to obtain a reaction mixed solution to be detected; and carrying out isothermal amplification and signal acquisition to obtain a detection conclusion. The multifunctional pretreatment buffer solution is used for cracking the to-be-detected sample, sample liquefaction, cracking and nucleic acid release are completed in a single system, the nucleic acid extraction and purification step of centrifugal column adsorption or magnetic bead elution is avoided, and the operation process is simplified.

The invention belongs to the technical field of plant fungus molecular biology detection, and discloses an LAMP primer group, a kit and a detection method for detecting oat smut pathogenic bacteria. A group of LAMP (loop-mediated isothermal amplification) specific primers are designed according to a specific sequence on a whole genome of the oat smut pathogenic bacteria Ustilago hordei, results are judged through a real-time fluorescence quantification method, an agarose gel electrophoresis method and an SYBR Green I fluorescent dye developing method, and the oat smut pathogenic bacteria carried by oat seeds are detected. The LAMP detection method for the oat smut pathogenic bacteria, established by the invention, is strong in specificity, high in sensitivity, high in speed and low in cost, provides a new technical means for detection of the oat smut pathogenic bacteria carried by the oat seeds, and has relatively high practical application value.

The invention discloses a method for detecting drug resistance of botrytis cinerea to succinate dehydrogenase (SDHI) bactericides. On the basis of an RPA-CRISPR / Cas12a system, a specific primer pair amplification target fragment and crRNA for activating Cas12a trans-cleavage activity are designed aiming at key mutation sites related to drug resistance in a bactericide action target SdhB gene, so that fluorescent light visible to naked eyes can be generated only when a resistant strain sequence is matched with the crRNA, and the fluorescent light is opposite to the fluorescent light generated when a sensitive strain sequence is matched with the crRNA. The method provided by the invention overcomes the dependence on laboratories and large instruments in the traditional drug resistance detection method, provides key technical support for accurate prevention and control and drug resistance management of grape gray mold, and provides direct drug use basis for scientific prevention and control in the field.

The invention belongs to the technical field of plant molecular breeding, and particularly relates to a KASP molecular marker related to the oil content of soybeans and application of the KASP molecular marker. The molecular marker is based on a soybean Glyma. 13G035200 gene, and targets an SNP (Single Nucleotide Polymorphism) site, which is remarkably associated with the soybean oil content, in the gene. The invention further provides a specific primer combination and a detection method of the molecular marker, soybean genotypes can be rapidly distinguished through PCR amplification and fluorescence signal detection, and high-oil-content soybean individuals can be accurately screened. The molecular marker has the advantages of being high in specificity, efficient in detection, reliable in result and the like, can be applied to soybean molecular marker assisted breeding, germplasm resource screening and high-oil soybean variety cultivation, remarkably shortens the breeding period, reduces the breeding cost, and provides an efficient tool for precise improvement of the soybean oil content character.

The invention relates to a primer probe combination and a kit for detecting eDNA of Chinese sturgeons and application of the primer probe combination and the kit. Specific primers and TaqMan probes are designed for a D-loop region of Chinese sturgeon mitochondrial DNA, and efficient specific amplification and absolute quantification of Chinese sturgeon eDNA in an environmental sample are realized by optimizing a reaction system and amplification conditions. Compared with the prior art, the method is simple and convenient to operate, does not need to perform harmful sampling on Chinese sturgeon individuals, is suitable for resource monitoring and protection evaluation in a large-range and dynamic environment, and has high sensitivity, high accuracy and high practical value.

The invention provides a KASP molecular marker related to mesosulfuron resistance of wheat and application of the KASP molecular marker, and belongs to the technical field of crop molecular genetic breeding. Through correlation analysis, it is found that the KASP molecular marker is remarkably correlated with mesosulfuron-methyl resistance, T / C polymorphism exists, and the mesosulfuron-methyl resistance of CC homozygous wheat is remarkably higher than that of TT homozygous wheat. On the basis, a KASP labeling reagent or kit containing a specific primer group is developed, the primer group comprises two allele specific upstream primers and a universal downstream primer, and the 5'ends of the upstream primers are connected with FAM and HEX fluorescent labels respectively. By optimizing a PCR reaction system and a landing procedure, high-throughput genotyping can be realized. The mesosulfuron-methyl-resistant wheat variety can be rapidly screened by applying the KASP molecular marker, a technical support is provided for herbicide resistance molecular breeding, the breeding efficiency of the resistant variety is remarkably improved, and the KASP molecular marker has important significance for guaranteeing safe production of wheat.

The invention relates to the technical field of molecular detection, discloses a KASP molecular marker related to a sheep backfat thickness character, a detection primer group and application of the KASP molecular marker and the detection primer group, and aims to provide a KASP molecular marker for detecting a sheep Chr15: g.12672188 site Tgt; the invention discloses a specific primer group for C variation, which consists of the following three primers: a site T allele typing primer FAM with a nucleotide sequence as shown in SEQ ID NO: 1, and a site T allele typing primer FAM with a nucleotide sequence as shown in SEQ ID NO: 2, the nucleotide sequence of the locus C allele typing primer VIC is as shown in SEQ ID NO: 2. The invention discovers and discloses Tgt of Chr15: g.12672188 site in a sheep genome for the first time; the C single nucleotide polymorphism (SNP) is obviously associated with the backfat thickness character. Association analysis of 118 Auodurau lake rams proves that the back fat thickness of CC and TT genotype individuals is obviously higher than that of TC genotype individuals. The discovery reveals that the two homozygous genotypes of the site are unique genetic patterns of dominant genotypes, and a direct and reliable molecular target is provided for genetic improvement of the sheep backfat thickness.

The invention discloses SSR (Simple Sequence Repeat) multiple PCR (Polymerase Chain Reaction) primers for megalobrama amblycephala paternity test and application, the primers comprise seven pairs of specific primers, namely a primer pair MamGLB2501, a primer pair MamGLB2502, a primer pair MamGLB2503, a primer pair MamGLG2504, a primer pair MamGLG2505, a primer pair MamGLR2506 and a primer pair MamGLR2507, and the base sequences of the seven pairs of specific primers are sequentially shown as SEQ ID NO.1-14; the invention also discloses an SSR multiplex fluorescent PCR method for megalobrama amblycephala paternity test by using the primer and application of the primer or the method in megalobrama amblycephala paternity test. The microsatellite site amplified by the primer is 3-6 base repetition, the typing is accurate, and the polymorphism of the amplified site is high; the method is accurate in identification and low in price.

The invention belongs to the technical field of bacterium detection, and discloses a multiple PCR primer for simultaneously detecting eight respiratory tract pathogenic bacteria and application thereof. On the basis of a Luminex liquid phase chip technology, specific primers are designed aiming at eight common respiratory tract infection pathogenic bacteria including pseudomonas aeruginosa, staphylococcus aureus, streptococcus pneumoniae, klebsiella pneumoniae, acinetobacter baumannii, stenotrophomonas maltophilia, haemophilus influenzae and escherichia coli; the multiple PCR primers which are high in amplification efficiency, good in specificity and suitable for simultaneously detecting the eight respiratory tract pathogenic bacteria are screened out, the multiple PCR system and microsphere hybridization conditions are continuously optimized, the multiple PCR method for the eight respiratory tract pathogenic bacteria is successfully constructed by applying the Luminex xTAG technology, and the multiple PCR method can be applied to rapid detection of multiple pathogenic microorganisms.

The invention relates to the technical field of bacterial detection, in particular to a nucleic acid colorimetric / photo-thermal dual-mode instant detection kit and system based on CRISPR / Cas12a and Au (at) PtNPs. One purpose of the invention is to provide a nucleic acid colorimetric / photo-thermal dual-mode instant detection kit based on CRISPR / Cas12a and Au (at) PtNPs. The nucleic acid colorimetric / photo-thermal dual-mode instant detection kit based on CRISPR / Cas12a and Au (at) PtNPs comprises the following components: Au (at) PtNPs coupled with Biotin-ssDNA-SH, a specific primer targeting a GBS conserved gene, a CRISPR / Cas12a cleavage reaction mixed solution and a reaction mixed solution. According to the technical scheme, the high robustness and reliability of the detection result in bedside, community clinics and other basic-level application scenes are jointly ensured through the simplified pretreatment process and the internal cross validation mechanism of the colorimetric / photo-thermal dual-mode signals, and a solid foundation is laid for wide popularization of the technology.

The invention relates to the technical field of biology, in particular to a primer probe combination product for respiratory tract infection detection. A plurality of specific primers and self-quenching probes are integrated in a single reaction system; simultaneous, rapid and rapid detection on 15 respiratory pathogens (including influenza A virus, influenza B virus, respiratory syncytial virus, severe acute respiratory syndrome coronavirus 2, mycoplasma pneumoniae, parainfluenza virus, rhinovirus, enterovirus, coronavirus, adenovirus, metapneumovirus, bordetella pertussis, bocavirus, chlamydia pneumoniae and chlamydia psittaci) is realized. The detection is accurate.

The invention belongs to the technical field of crop molecular markers, and particularly discloses a molecular marker for identifying the salt tolerance of brassica napus in the germination period and application of the molecular marker. The molecular marker is located at the position of an A02 chromosome scafoldA028707654 of a brassica napus reference genome ZS11.v10, and the allelotype of the molecular marker is wild type C or mutant type CATATT insertion. The invention also provides a specific primer group for detecting the molecular marker, and the salt tolerance (sensitive type or tolerance type) of the brassica napus line can be quickly identified according to the existence of 411bp and 416bp bands through double PCR amplification and electrophoresis detection. The molecular marker is significantly associated with salt tolerance phenotypes, the detection method is rapid, accurate and low in cost, the molecular marker can be used for brassica napus salt tolerance germplasm resource screening and molecular marker-assisted breeding, and the breeding efficiency is significantly improved.

The application relates to a sarcoma fusion gene nanopore sequencing primer set and application thereof, wherein the primer set comprises five pairs of specific primers (Primer 1-5) with fixed sequences, each primer follows the design principle of 18-25 bp in length, 40%-60% in GC content and strict matching of a target sequence at a 3' end, and the length of an amplification product is 300-800 bp, which is suitable for nanopore sequencing. Through qPCR verification, the primer set has single-peak melting curves, close-to-100% amplification efficiency, strong specificity and excellent amplification performance.

The application relates to a primer probe composition, a kit and a detection method for specifically detecting Blattella germanica. The specific primer and TaqMan probe are designed and screened based on the specific sequence of the mitochondrial COI gene of Blattella germanica. The detection method has extremely high specificity, has no cross reaction with close species such as Periplaneta americana and Blattella germanica, has high sensitivity, the minimum detection limit is 1 fg / muL, and has good repeatability, with the variation coefficients of groups and between groups being less than 3%.

The invention provides a method and a kit for quantitative detection of recombinant redistribution of a Databa virus based on a multiple digital PCR (Polymerase Chain Reaction) technology, and belongs to the field of infectious disease monitoring. The method comprises the following steps: providing a to-be-detected sample containing segmented viruses and viral nucleic acid; detecting nucleic acid by using a plurality of multiple digital PCR reaction systems containing specific primer pairs and fluorescence labeled probes; amplifying according to a fluorescence coding rule, collecting and analyzing multi-channel fluorescence signals, and calculating the absolute copy number of each genotype; and based on copy number analysis genotype intra-fragment distribution proportion and inter-fragment combination mode, evaluating characteristics and frequency of virus recombination reconfiguration events. According to the detection method and the kit provided by the invention, accurate quantification of all main genotypes of the Databa virus can be realized.

The application belongs to the technical field of molecular biology, discloses a SNP molecular marker closely linked with the genetic sex of Ophiocephalus argus and application thereof, and specifically discloses the SNP molecular marker closely linked with the genetic sex of Ophiocephalus argus, the sequence of the SNP molecular marker is shown as SEQ ID NO:1, the SNP site is located at the 151th position from the 5' end of the sequence shown as SEQ ID NO:1, and the base is T / G. The application provides a SNP molecular marker closely linked with the sex gene of Ophiocephalus argus, and the genetic sex of Ophiocephalus argus can be accurately identified through the single base difference in the molecular marker. A specific KASP primer for amplifying the molecular marker closely linked with the sex gene of Ophiocephalus argus is also provided, the genetic sex of Ophiocephalus argus can be detected, the method is simple and easy to use, has high analysis flux and high accuracy, and is suitable for the detection of a large number of samples.

The invention discloses a specific primer group, a kit and a method for detecting double photosynthetic bacteria, and relates to the technical field of microbiological detection. The specific primer group is designed based on a conserved region of a photosynthetic reaction center L subunit gene (pufL) of rhodopseudomonas and ectothiorhodospirillum, the conservative property is greater than or equal to 99%, the similarity rate is greater than or equal to 98.5%, specific amplification of target bacteria can be realized, and cross amplification interference is avoided. The matched kit comprises the primer group, 2 * SYBR Green qPCR Master Mix, a positive control component, a negative control component and the like, and the detection method realizes rapid and sensitive detection of the two photosynthetic bacteria by optimizing sample pretreatment, PCR amplification conditions and result interpretation standards. The detection specificity is high, the sensitivity is high (the lowest detection limit reaches 10 < 2 > cfu / mL level), the detection period is short (1-2 hours), the applicability is wide, the method can be used for detecting various samples such as river water, aquaculture water and microbial inoculum fermentation liquor, one set of primers can detect two target bacteria at the same time, the detection cost and the operation complexity are reduced, and the method is suitable for aquaculture environment monitoring and microbial inoculum application quality control.

The invention provides a capture primer group and a kit for targeted Salivirus genome sequencing and application of the capture primer group and the kit, and belongs to the technical field of genome sequencing. The targeted capture primer group of the Salivirus virus genome comprises a primer 1 and a primer 2, the nucleotide sequence of the primer 1 is as shown in SEQ ID NO. 1, and the nucleotide sequence of the primer 2 is as shown in SEQ ID NO. 2. According to the invention, a universal reverse transcription oligo-T primer and the Salivirus specific primer group provided by the invention are put into the reverse transcription step at the same time, so that not only is a macro transcription effect achieved, but also a Salivirus sequence in a sample is synchronously identified for specific capture. According to the method, the complete Salivirus genome sequence can be efficiently obtained, and the method has good coverage and sequencing adaptability to the virus genome.

The invention discloses RT-qPCR-HRM (real-time quantitative polymerase chain reaction-high-resolution melting) detection primers and a detection method for identifying five different subtype avian influenza viruses, and belongs to the technical field of biology. The invention discloses an RT-qPCR-HRM detection method for identifying five different subtypes of avian influenza viruses, which comprises the following steps: designing a specific primer based on a highly conserved region of an AIV M gene, and simultaneously carrying out genetic typing on five subtypes of AIV strains H3, H5, H6, H7 and H9 in combination with an HRM technology; by comparing melting curve characteristics of M gene segments of different subtypes of AIV, an AIV detection method with good sensitivity and specificity is established. According to the method, the five AIVs of H3, H5, H6, H7 and H9 can be accurately typed, the defect of missing detection of a traditional AIV detection method is overcome, and the method has a wide application prospect.

The application provides a gene, a primer set and an application for identifying golden ear, and belongs to the technical field of biological detection. The application obtains specific gene fragments of two species contained in the golden ear through large-scale whole genome comparison analysis, designs species-specific primers based on the specific gene fragments, and further realizes identification of the effective species of the golden ear. The identification is extremely crucial in the production process of the effective species of the golden ear, and can maximize the guarantee of the fruiting rate in the production of the golden ear.

The invention relates to the technical field of plant molecular breeding, in particular to a Kasp marker EL-5-KASP-387 closely linked with the ear length of corn and application of the Kasp marker EL-5-KASP-387, an Indel functional site corresponding to the Kasp marker is located at the position 184049387 of the fifth chromosome of corn, the polymorphism is- / TGA, and a matched specific primer group is composed of two forward primers and a reverse primer. The major site of the ear length is located through whole genome association analysis, the developed KASP marker can accurately distinguish three genotypes, ear length character identification can be completed in the seedling stage, and the multi-environment weighted matching degree of the genotypes and phenotypes reaches 99.2%. The method does not need gel electrophoresis, has the advantages of high flux and high precision, can greatly shorten the breeding period of the long-ear corn, improves the breeding selection accuracy, and provides efficient technical support for high-yield molecular directional breeding of the corn.

The present application relates to the technical field of molecular detection, and particularly relates to a specific primer combination for distinguishing Enteromorpha prolifera and related species, application and molecular identification method thereof. The primer combination comprises an upstream primer SEQ1 and a downstream primer SEQ2. The primer sequence of SEQ1 is shown as SEQ ID NO. 01, and the primer sequence of SEQ2 is shown as SEQ ID NO. 02. The primer combination of the present application is designed based on the genomic difference of two kinds of algae, has high specificity and sensitivity, can effectively avoid cross reaction, and ensures the accuracy of the detection result. Meanwhile, combined with the real-time fluorescent quantitative PCR (qPCR) technology, the method can effectively quantify the abundance of Enteromorpha prolifera in environmental samples or laboratory samples, and provides reliable technical support for early warning, tracing analysis and prevention and control strategy of green tide outbreak.

The invention belongs to the technical field of biology, and relates to a dual RPA (recombinase polymerase amplification) fluorescent rapid detection method for simultaneously detecting Globoderma rostochiensis and Meloidogyne enterolobii. According to the characteristics of ITS sequences, obtained through amplification, of the potato golden nematode and the elephant bean root-knot nematode, specific primers GrF / GrR and Me2F / Me2R and two fluorescence detection probes GrPe and MePe of the potato golden nematode and the elephant bean root-knot nematode are respectively designed on the basis of specific fragment sequences, a dual RPA reaction system is established, DNA extraction is carried out on the potato golden nematode and the elephant bean root-knot nematode, and the specific primers GrF / GrR and Me2F / Me2R and the two fluorescence detection probes GrPe and MePe of the elephant bean root-knot nematode are obtained. A real-time fluorescent PCR detector can be used for detecting the potato golden nematode and the elephant trunk bean root-knot nematode at the same time. According to the detection method, an RPA-isothermal amplification fluorescence detection method is high in specificity, high in sensitivity and simple and convenient to operate.

The invention discloses a method and a device for detecting CAR (Chimeric Antigen Receptor) cells. The method comprises the following steps: taking a genome of a sample to be detected as a template, carrying out fluorescent quantitative PCR reaction by using a primer and a probe aiming at the sequence of a CD8 alpha transmembrane region-41 BB costimulatory molecule in a CAR gene to obtain a CT value, and calculating the copy number of the CAR gene according to the CT value and a standard curve to obtain the CAR cell distribution. Specific primers and probes are designed for CAR cells containing CD8 alpha transmembrane region-41BB costimulatory molecules, a fluorescent quantitative PCR method for detecting the copy number of CAR genes is developed, then the CAR cells are quantified, and it is proved that the method has wide applicability through multi-angle verification analysis and limitation of detection standards.

The invention belongs to the technical field of molecular markers, and particularly relates to an SNP molecular marker for high-quality protein waxy corn and application of the SNP molecular marker. The SNP molecular marker is closely linked with functional mutation of a high-quality protein corn o2 gene, and a non-synonymous mutation site base of the SNP molecular marker site is T / C and is located on a corn gene Zm00001d018971. According to the SNP molecular marker developed on the basis of o2 gene mutation sites, Zm00001d018971 variant and wild type alleles can be distinguished specifically and highly sensitively, and a pair of specific primer groups capable of amplifying the SNP molecular marker is provided. The o2 mutant corn sample can be quickly, efficiently and accurately identified only by performing PCR amplification on the corn plant by using the specific primer group disclosed by the invention, lossless and accurate genotype identification in the seedling stage is realized, and reliable and low-cost technical support can be provided for efficient and large-scale breeding of high-quality protein corn.

The invention provides a ready-to-use PCR tube preloaded with Taq polymerase and reaction components and stable at room temperature, and belongs to the field of molecular biology and nucleic acid diagnosis. The invention provides a ready-to-use PCR (polymerase chain reaction) tube which can be stored at room temperature and is pre-filled with a premix for carrying out polymerase chain reaction (PCR) amplification on all necessary components in advance. The kit is characterized in that each tube is pre-filled with locally prepared Taq polymerase, a PCR buffer solution, dNTPs, MgCl and target specific primers (only a customized PCR kit-ready-to-use PCR plus tube contains the primers), and all the components are dried and stored in a dry powder form, so that the shelf life can be prolonged, and the tedious steps of on-ice operation can be avoided. According to the method, the error rate in the PCR preparation process is reduced to the maximum extent, and the diagnosis and research work flow is simplified.