43results about How to "Primer specificity is high" patented technology

The invention discloses a universal RT-PCR (Reverse Transcription Polymerase Chain Reaction) detection primer and a detection method for avian pneumovirus (APV), belonging to the technical field of detection for avian viruses.. The universal RT-PCR detection primer has nucleotide sequences shown as SEQ ID NO:1 to SEQ ID NO:2. The detection method is as follows: the universal RT-PCR detection primer for avian pneumovirus and the RNA (Ribonucleic Acid) of a sample to be detected are subjected to RT-PCR, and the presence of avian pneumovirus in the sample to be detected is confirmed if the PCR product fragment is 319bp o 320bp. The method disclosed by the invention is high in primer specificity, small in fragment and high in sensitivity, can finish the entire PCR process within 2 hours and can also specifically detect the presence of various subtype APVs, thus greatly reducing the erroneous diagnosis for APV infection.

The invention provides a method and a system for realizing Y-STR typing on the male individual by adopting 26 Y-STR genetic loci. The method comprises the following steps: acquiring DNA of the male individual, and acquiring the genotypes of the 26 Y-STR genetic loci of the DNA, wherein the 26 Y-STR genetic loci are as follows: DYS19, DYS385a / b, DYS389I / II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS448, DYS456, DYS458, DYS635, GATA H4, DYS438, DYS439, DYS460, DYS447, DYS527a / b, DYS617, DYS522, DYS444, DYS508 and DYS533; and acquiring the Y-STR typing result of the male individual according to the genotypes of the 26 genetic loci of the male individual. With the typing result, the operations including family checking, paternity identification, detection for the male element in male and female mixed samples, and the detection for the male element in a plurality of male mixed samples can be realized on the male individual.

The invention relates to the technical field of security detection of agricultural and sideline products, in particular to a primer for simultaneously detecting five food-borne pathogenic bacteria through a liquid chip and a detection method. The detection method comprises steps as follows: a specific primer and a gene probe for five foodborne pathogenic bacteria are designed; DNA templates of the detected five foodborne pathogenic bacteria are prepared; PCR (polymerase chain reaction) amplification is performed on the five foodborne pathogenic bacteria; coupling is performed on the gene probe for the five foodborne pathogenic bacteria and microspheres, and a microsphere probe is obtained and verified; an obtained PCR amplification product and the microsphere probe have a hybridization reaction; a fluorescence signal of a hybridized product is detected, and quantitative analysis is performed on the five foodborne pathogenic bacteria. A method capable of simultaneously detecting five common foodborne pathogenic bacteria is successfully established, the variable coefficient is within 2%, the specificity is 100%, the sensitivity can reach 100 CFU / mL, and the method is equivalent to a fluorescent PCR technology and can be applied to high-throughput rapid detection of clinic and environment specimens of foodborne diseases.

The invention discloses a method and a kit for detecting mutation of an EGFR gene. The method comprises the steps of bonding a specific primer with a target template DNA sequence, amplifying a to-be-tested sample target region by virtue of a universal primer, purifying a library by virtue of magnetic beads, carrying out high-throughput sequencing on the obtained library, and analyzing the mutation of the EGFR gene. The kit comprises the primers, a PCR mixing reagent, a positive control and the magnetic beads. By utilizing the method and the kit, the experimental steps can be simplified, and the detection sensitivity is improved. The method and the kit are applicable to the rapid detection of the mutation of the EGFR gene of clinic tumor patients and are helpful for guiding clinic personalized medication.

The invention provides method and system for Y-STR typing of an individual man. The method comprises the following steps: acquiring the DNAs of the individual man, wherein acquired DNAs comprise genotypes of 25 gene loci including 24 Y-STR gene loci and a gender determination locus Amelogenin, and the 24 Y-STR gene loci are DYS460, DYS389I / II, DYS390, DYS392, DYS458, DYS437, DYS385a / b, GATA_H4, DYS522, DYS456, DYS391, DYS447, DYS438, DYS448, DYS393, DYS635, DYS439, DYS19, DYS444, DYS527a / b, DYS617; and acquiring the Y-STR typing results of the individual man according to the 25 gene loci of the individual man. The Y-STR typing results can be used for family checking, male paternity testing, detection of male components in a hybrid male and female sample, detection of male components in a mixed sample of a plurality of males and the like for the individual man.

The invention discloses primers for identifying tapisicia sinensis gender and application thereof. The primers include TSMHF and TSMHR, wherein the sequence of TSMHF is 5'-TTGTCCCTCTCAACTTCGCT-3', and the sequence of TSMHR is 5'-AAAATCAACCAGCCAGTTCG-3'. The primers are used for identifying male and bisexual tapisicia sinensis. The primers have the advantages that one pair of specific microsatellite marker primers for identifying tapisicia sinensis gender is obtained by accident from 150 pairs of microsatellite primers, the primers are clear in banding pattern, good in repeatability, capable of identifying tapisicia sinensis gender with an identification rate of 100%, high in reliability, and capable of being used for identifying tapisicia sinensis gender; the primers are extremely high in specificity and stability, only the steps of conventional PCR, polyacrylamide gel electrophoresis and dye development are needed, fragment separation is not needed, tapisicia sinensis gender can be identified by naked eye observation, and theoretical and technical guidance is provided to the directive breeding of tapisicia sinensis.

The invention provides a method and system for Y-SNP typing of male individuals. The method comprises the following steps: acquiring the DNAs of a male individual; acquiring the genotypes of ten Y-SNPsites of the DNAs, wherein the ten Y-SNP sites include M130, M174, M89, M201, M304, M9, M231, M175, M242 and M207; and acquiring the Y-SNP typing results of the male individual according to the ten sites of the male individual. The invention also provides a system for rapid Y-SNP typing of male individuals. The method and system provided by the invention can accurately acquire the genotypes of ten Y-SNP sites of the male individual, so the Y-SNP typing results of the male individual can be acquired; and the Y-SNP typing results can provide data support for family checking, male paternity identification, or ethnic derivation deduction of the individual man.

The invention provides an SNP marker related to a pepper tomato spotted wilt virus disease resistance gene as well as specific primers and application of the SNP marker. A nucleotide sequence of the SNP marker related to the pepper and tomato spotted wilt virus disease resistance gene is as shown in SEQ ID NO.1, and the SNP marker is a 21st basic group of the sequence as shown in SEQ ID NO.1. Theinvention also provides the specific primers and a corresponding test kit for identifying the SNP marker, and provides an identification method for pepper resisting the tomato spotted wilt virus disease by using the specific primers or the test kit. By adopting the SNP molecular marker disclosed by the invention, the pepper TSWV (Tomato Spotted Wilt Virus) disease resistance gene can be quickly and accurately judged only by carrying out DNA detection on the pepper plant in a seedling stage without artificial inoculation identification of pepper TSWV disease resistance, and large-batch detection can be carried out on samples, so that the breeding efficiency is improved, and the identification time is shortened.

The invention discloses a PCR (Polymerase Chain Reaction) detection primer and method for H120 strain of avian infectious bronchitis virus, belonging to the technical field of detection of avian virus. The PCR detection primer comprises two pairs of detection primers, wherein the nucleotide sequences are respectively SEQIDNO: 1-2 and SEQIDNO: 3-4. By use of the two pairs of primers, RNA (Riboneucleic Acid) of a sample to be detected is used as a template so as to carry out RT-PCR amplification, and if products of two types such as 653bp and 230bp are amplified simultaneously, the containing of the H120 of the IBV in the sample to be detected can be judged. The PCR detection primer has the advantages that the specificity of the primer is high, the segments are small, the whole RT-PCR process can be finished in 2-2.5 hours, the H120 vaccine strain of the IBV can be quickly identified without the sequencing need of the sequencing company, so that the time is greatly saved for clinic detection.

The invention discloses a DNA examination reagent for sample examination, and a special primer thereof. A DNA examination primer group provided by the invention is composed of a primer pair group used for amplifying 21 Y-STR sites, a primer pair for amplifying a sex site Amelogenin, and a primer pair for amplifying an autosome SRT site. Experiments prove that the DNA examination reagent can simultaneously examine the 21 Y-STR sites and the autosome SRT site, and the primer specificity is high, so complete SRT typing can be obtained, peak patterns are sharp and intelligible, the balance is good, no Pull-up peaks or stutter bands appear, no non-specific artificial products appear, and legal medical experts' DNA large scale sample examination requirements are completely met.

The invention discloses a follicle-stimulating hormone receptor activity detection kit and a detection method thereof. The detection kit comprises two pairs of amplification primers and a multiplex PCR mixed reaction solution, wherein the amplification primers are used for amplifying obese gene polymorphic sites and comprise rs6165 sites and rs6166 sites, and each pair of primers respectively corresponds to upstream and downstream regions of one SNP site. According to the follicle-stimulating hormone receptor activity detection kit and the detection method thereof, a multiplex PCR amplification method is adopted, two important SNP sites having high relevancy with FSHR activity are amplified at the same time, the amplified site combination can be flexibly selected according to actual needsto achieve the function of one kit with multiple functions, and primers in the kit are high in specificity and low in mismatch rate. The primers are reasonable in length and Tm value design, and the amplified product is convenient to detect and isolate. Multiple target products are amplified once, so that the amplification efficiency can be greatly improved, and the amplification cost can be reduced. The detection kit can be extended to other SNP site detection, and has a good popularization value.

The invention discloses a detection method for EV71 virus in an environmental water body, wherein a semi-nested RT-PCR (Reverse Transcription-Polymerase Chain Reaction) and real-time fluorescence quantification RT-PCR reaction system is established to detect EV71 virus in the environmental water body by using a recombinant plasmid as a reference product, and the recombinant plasmid is prepared by using a primer and a probe aiming at EV71 virus designed according to a genetic conservative fragment of enterovirus belonging to a VP1 serotype area. The method has good specificity and high sensitivity and precision, and can qualitatively and quantitatively detect EV71 virus in the environmental water body accurately.

The invention discloses a detection method for CVA16 in an environmental water body, wherein a semi-nested RT-PCR (Reverse Transcription-Polymerase Chain Reaction) and real-time fluorescence quantification RT-PCR reaction system is established to detect CVA16 virus in the environmental water body by using a recombinant plasmid as a reference product, and the recombinant plasmid is prepared by using a primer and a probe aiming at CVA16 virus designed according to a genetic conservative fragment of enterovirus belonging to a VP1 serotype area. The method has good specificity and high sensitivity and precision, and can qualitatively and quantitatively detect CVA16 virus in the environmental water body accurately.

The invention discloses a quick amplifying reagent kit and amplifying method of a CYP2C19 gene. The reagent kit comprises 3 pairs of amplifying primers, Taq enzymes, dNTPs and a buffered solution, wherein the amplifying primers are used for amplifying at least two SNP sites in a CYP2C19*2 type, a CYP2C19*3 type and a CYP2C19*17 type, and each pair of primers corresponds to upstream and upstream regions of one of SNP sites. A multiplex-PCR amplifying method is used, a plurality of important SNP sites of the CYP2C19 are amplified at the same time, the combination of sites to be amplified can beflexibly selected according to actual demands, and the effect that one reagent kit has many functions can be achieved; the primers in the reagent kit are high in specificity and low in mispairing rate, the length and the Tm value of the primers are reasonable to design, and amplified products are convenient to detect and separate. Multiple purpose products are amplified at a time, so that the amplifying efficiency can be greatly improved, and the amplifying cost can be reduced.

The invention belongs to the technical field of biology, and provides a special primer for simultaneously and accurately detecting SNP loci of related genes of 'slap-induced deafness' and ' injection-induced deafness'. The special primer comprises a primer group 1 and a primer group 2, wherein the primer group 1 is a specific PCR primer composed of 13 pairs of primers, and the primer group 2 is asingle-base extension primer composed of 13 primers. The invention further provides a preparation for simultaneously and accurately detecting the SNP loci of the related genes of 'slap-induced deafness' and 'injection-induced deafness', and a method for simultaneously and accurately detecting the SNP loci of the related genes of 'slap-induced deafness' and 'injection-induced deafness'. By adoptingthe special primer provided by the invention, accurate prediction of high-risk groups of 'slap-induced deafness' and 'injection-induced deafness' can be efficiently completed simultaneously through one-time detection, the detection time is greatly shortened, the cost is saved, and the special primer has very high application value for preventing 'slap-induced deafness' and 'injection-induced deafness' and 'deafness-induced dumbness' caused by 'slap-induced deafness' and 'injection-induced deafness'.

The invention provides a molecular marker of a rice brown planthopper-resistant gene BPH39 and an application thereof. A local high-density physical map covering the BPH39 is constructed by constructing a near-isogenic line of the BPH39 and analyzing a recombinant exchange single plant of a high-generation backcross population, and the gene is finely positioned in an interval of 30kb. The two molecular markers ZL1607 and ZL2137 in the interval are closest to the BPH39, and the reliability of a single plant for screening a target gene by utilizing the two markers is the highest. Besides, othermolecular markers, I1540, I32-4 and ZL6061 which are closely linked with the gene can be used for screening single plants of the target gene, so that the selection efficiency of brown planthopper-resistant rice varieties or strains can be greatly improved, and the breeding age limit is shortened.

The invention discloses a method for fast identifying the generation of an EC (ethyl carbamate) precursor of citrulline from lactic acid bacteria, and belongs to the technical field of microbiological detection. By the method, whether a lactic acid bacterium strain has an arginine deiminase path or not and whether citrulline can be formed by using arginine or not can be judged, so that whether the lactic acid bacterium strain in the fermented food and beverage has the possibility to generate a cancerogen of EC or not can be determined. The method disclosed by the invention is fast, simple, convenient and accurate; the degenerate primer specificity is high; the universality is high; the lactic acid bacteria of different species from different food can be detected; good food safety detection prospects are realized.

The invention relates to the technical field of rice breeding and molecular biology, in particular to a major QTL for regulating and controlling the brown rice rate of rice, a molecular marker and application. The invention discloses a major QTL for regulating and controlling the brown rice rate of rice. The major QTL is located on a chromosome 10 of the rice and is named as QBRR-1, the genetic distance is 70.1-81.73 cM, and the physical distance is 17570591-19066686 bp. The invention also provides the molecular marker of the major QTL for regulating and controlling the brown rice rate of the rice. According to the major QTL, the rice with high brown rice rate is bred through the molecular marker.

The invention relates to a method and kit for identifying genuine medicinal materials herba cynomorii different in growing area. The method comprises the steps that firstly, genome DNA of a to-be-detected sample serves as a template, and the segment containing the sequence shown in SEQ ID NO.1 is amplified; secondly, an amplified product is sequenced, after splicing, a primer area is deleted so as to obtain a complete amplified segment containing the sequence shown in SEQ ID NO.1, and basic groups at the 188th position and the 304th or 305th position starting from the 5' terminal in the sequence shown in SEQ ID NO.1 are detected; if the basic group at the 188th position starting from the 5' terminal in the sequence shown in SEQ ID NO.1 is T, or the basic group at the 304th position is A, or the basic group at the 305th position is C, the herba cynomorii is identified as Tacheng herba cynomorii, and if the basic group at the 188th position starting from the 5' terminal in the sequence shown in SEQ ID NO.1 is C, or the basic group at the 304th position is T, or the basic group at the 305th position is T, the herba cynomorii is identified as herba cynomorii from other growing areas. The method is wider in adaptability, high in primer specificity, high in amplification efficiency and capable of detecting any samples of herba cynomorii from which DNA can be extracted.

The invention relates to the technical field of molecular biology detection, in particular to a RT-PCR detection primer and a detection method for K subgroup avian leukosis virus. The primers of the present invention are represented by SEQ ID NO:1 and SEQ ID NO:2. The RT-PCR detection method of K subgroup avian leukosis virus of the present invention comprises the following operations: (1) RNA template preparation; (2) amplifying the target fragment; (3) identifying the amplified product by electrophoresis. The primers of the present invention have high specificity and short extension fragments, and the entire PCR identification process can be completed within 2 to 2.5 hours, and the K subgroup avian leukosis virus can be quickly identified without sequencing. The diagnosis saves a lot of time. The method of the invention is simple to operate, the equipment used in ordinary experiments is easy to configure, the detection cost is low, and it is convenient for clinical popularization and application.

The invention belongs to the technical field of rice molecular marker preparation, and in particular relates to the molecular markers of two major brown planthopper-resistant rice genes QBph3 and QBph4 derived from the insect-resistant introduction line IR02W101 at the same time. The genotypes of each individual plant of BC1F2 were crossed, combined with the resistance levels of N. lugens planthoppers at the seedling stage of each F2:3 family for genetic linkage analysis, the resistance gene QBph3 of IR02W101 was finely mapped between the markers t6 and f3 on the long arm of the third chromosome , obtained co-segregation marker c3-14, closely linked markers q1 and m3; QBph4 was fine-mapped between markers p17 and xc4-27 on the short arm of chromosome 4, and closely linked markers p6, p9, c4-5, xc4-7 were obtained , HJ16, J417 and IN156. The invention can effectively detect whether the main effect gene locus is contained in the insect-resistant introduction line IR02W101 and derivative varieties.

The invention discloses a detection method for CVA16 in an environmental water body, wherein a semi-nested RT-PCR (Reverse Transcription-Polymerase Chain Reaction) and real-time fluorescence quantification RT-PCR reaction system is established to detect CVA16 virus in the environmental water body by using a recombinant plasmid as a reference product, and the recombinant plasmid is prepared by using a primer and a probe aiming at CVA16 virus designed according to a genetic conservative fragment of enterovirus belonging to a VP1 serotype area. The method has good specificity and high sensitivity and precision, and can qualitatively and quantitatively detect CVA16 virus in the environmental water body accurately.

The invention discloses a primer for diagnosing mulberry blight. The base sequence of the upstream primer is shown in SEQ ID NO: 1, and the base sequence of the downstream primer is shown in SEQ ID NO: 2. The invention also discloses a method for diagnosing bacterial wilt by using the primer, which comprises the following steps: peeling and chopping the mulberry roots sterilized on the surface, placing them in sterile water, and centrifugally oscillating to prepare an aqueous suspension, and The aqueous suspension is used as a template, and the primers are used to carry out PCR amplification, and the amplified product is separated by gel electrophoresis and developed with an indicator. If there is a DNA band on the gel, the mulberry tree is infected with bacterial wilt, such as If there is no DNA band, the mulberry tree is not infected with bacterial wilt. The invented primer has high specificity, and the detection period of the mulberry bacterial wilt is short by using the primer, which saves more than 90% of the time compared with the conventional method, and has high sensitivity and accuracy.

The invention belongs to the technical field of biology, and provides a special primer for detecting SNP sites of invasive aspergillosis risk after allogeneic hematopoietic stem cell transplantation.The special primer comprises a first primer group and a second primer group; the first primer group is a specific PCR primer consisting of three pairs of primer groups; and the second primer group isa single base extension primer consisting of three primers. The invention also provides a preparation of the SNP sites for detecting the invasive aspergillosis risk after allogeneic hematopoietic stemcell transplantation, and an SNP sites information method for detecting the invasive aspergillosis risk after allogeneic hematopoietic stem cell transplantation. By adopting the special primer for detecting the SNP sites of the invasive aspergillosis risk after allogeneic hematopoietic stem cell transplantation provided, the correlation between the gene polymorphism of the three SNP sites relatedto the attack risk of invasive aspergillosis after allogeneic hematopoietic stem cell transplantation and the occurrence risk of invasive aspergillosis after allogeneic hematopoietic stem cell transplantation can be efficiently and simultaneously completed through one-time detection, so that the detection time is greatly shortened, and the cost is saved.

The invention provides a SNP marker related to the resistance gene of pepper tomato spotted wilt virus disease and its specific primer and application, and the nucleotide sequence of the SNP marker related to the resistance gene of pepper tomato spotted wilt virus disease is as shown in SEQ ID As shown in NO.1, the SNP marker is the 21st base of the sequence shown in SEQ ID NO.1. The present invention also provides specific primers and corresponding kits for identifying the SNP marker, and provides a method for identifying peppers against tomato spotted wilt virus disease using the specific primers or kits. Using the SNP molecular marker of the present invention, the TSWV disease resistance gene of pepper can be quickly and accurately judged without the need for artificial inoculation identification of pepper TSWV disease resistance. Samples are tested in large quantities to improve breeding efficiency and shorten identification time.

The invention relates to the molecular biological technology field, and concretely relates to an SSR primer group and a method for identification of hot pepper species thereby. The SSR primer group is composed of 20 primers. The method for identification comprises steps: first, DNA extraction of hot pepper leaves; second, PCR amplification; third, electrophoresis detection; fourth, data recording; and fifth, result determination. The primers have high specificity and high amplification efficiency, and can identify different species of hot peppers rapidly. The method is advantaged by short period, low cost, reliable results, manpower saving, material resource saving and land resource saving.

The invention discloses a detection method for EV71 virus in an environmental water body, wherein a semi-nested RT-PCR (Reverse Transcription-Polymerase Chain Reaction) and real-time fluorescence quantification RT-PCR reaction system is established to detect EV71 virus in the environmental water body by using a recombinant plasmid as a reference product, and the recombinant plasmid is prepared by using a primer and a probe aiming at EV71 virus designed according to a genetic conservative fragment of enterovirus belonging to a VP1 serotype area. The method has good specificity and high sensitivity and precision, and can qualitatively and quantitatively detect EV71 virus in the environmental water body accurately.

The invention discloses a DNA testing reagent and special primers for sample screening. The present invention provides a DNA test primer set, consisting of a primer pair set for amplifying the following 21 Y-STR sites, a primer pair for amplifying sex locus Amelogenin, and a primer for amplifying autosomal STR sites pair composition. Experiments of the present invention prove that the DNA detection reagent provided by the present invention can simultaneously check 21 Y-STR loci and 1 autosomal STR locus, and the primers have high specificity, can obtain complete STR typing, and peak type Sharp and clear, well-balanced, no Pull-up peaks, stutter bands, and non-specific artifacts, it can fully meet the requirements of large-scale forensic DNA sample testing.