37results about How to "Accurate amplification" patented technology

The invention relates to a scophthalmus maximus T170G single nucleotide polymorphic marking detection method. The method comprises the following steps: extracting scophthalmus maximus genome DNA and diluting for later use; analyzing the sequences of EST, screening the sequence containing a candidate SNP locus, designing a specific primer at the two ends and designing a non-marking probe with the closed 3' end in front of and behind the locus (comprising the locus); performing asymmetric PCR amplification on the scophthalmus maximus group genome DNA by using the primer; hybridizing the amplification product with the non-marking probe with the closed 3' end; and placing the hybrid product on a LightScanner, and detecting and analyzing the melting curve to obtain the genetic polymorphism mapof the scophthalmus maximus. The method is applicable to detection technologies of scophthalmus maximus genetic marking, genealogy authentication, genetic linkage map construction and the like. According to the method which is convenient, quick and accurate, the scophthalmus maximus T170GSNP marked genetic variation map can be obtained quickly; and the genotype of each individual of the scophthalmus maximus can be detected intuitively.

The present invention relates to a method for accurately detecting a target nucleic acid by asymmetrically and isothermally amplifying the target nucleic acid using an external primer set, an internal primer set having different percentages of forward and reverse DNA-RAN-DNA hybrid primers, and a DNA-RNA-DNA hybrid signal probe and amplifying a signal of the probe at the same time. According to the present invention, the signal of the probe can be efficiently amplified compared with the method of the prior art, a symmetric iTPA method, which is an isothermal primer and probe amplification method using the same percentage of primers. Therefore, the present invention is applied to the accurate detection and confirmation of pathogens, detection of gene modification inducing identified phenotypes, diagnosis of susceptibility to genetic diseases, evaluation of gene expression, and various genome projects, and thus is useful in the molecular biological researches and disease diagnosis.

The invention discloses a method for detecting C135T single nucleotide polymorphism (SNP) of turbot. The method comprises the following steps of: extracting genome DNA of turbot and diluting for later use; analyzing a sequence of an expressed sequence tag (EST), screening a sequence containing a candidate SNP locus, designing specific primers at two ends of the sequence, and designing 3' end-closed unlabeled probes before and after the locus (including the locus); performing asymmetrical polymerase chain reaction (PCR) amplification on population genome DNA of turbot by using the primers; hybridizing an amplification product and the 3' end-closed unlabeled probes; and detecting a melting curve of the hybrid on a light scanner, and analyzing the melting curve to obtain a genetic polymorphism map of turbot. The method is suitable for detection technologies such as turbot population genetic markers, lineage authentication, genetic linkage map construction and the like; and by the method,a genetic variation map of the C135TSNP of turbot can be quickly, easily and accurately obtained, and the genotype of each individual of turbot can be intuitively detected.

The invention belongs to the field of pathogen detection, and provides a primer pair for quantitatively detecting a yellow leaf curl virus, application of the primer pair to quantitative detection of the yellow leaf curl virus and a method for quantitatively detecting the yellow leaf curl virus. The primer pair for quantitatively detecting the yellow leaf curl virus can be used for efficiently and accurately amplifying a target fragment. The method for quantitatively detecting the yellow leaf curl virus by virtue of the primer pair can be used for accurately and rapidly identifying pathogenic bacteria and accurately quantifying a viable count.

The invention discloses a primer group capable of accurately measuring the ITS base sequence of cerasus schneideriana, synthesis and rapid molecular identification. The sequence of the primer group isshown as follows: 1f:5'-CTCCTCGTCCCTTTTCTC-3' (SEQ ID No.1) and 2r:5'-AGTTTCTTTTCCTCCGCT-3' (SEQ ID No.2). The invention relates to a specific pair of primers of 1f-2r, the primers can be used to acquire the ITS sequence of the cerasus schneideriana with good quality through PCR, and the 1f-2r is specific sequence primers for the cerasus schneideriana and used for ITS molecular sequence auxiliaryidentification by combining morphological classification; and in addition, the specific primer pair of 1f to 2r can accurately amplify the ITS sequence of the cerasus schneideriana and can provide acertain basis for population genetic diversity, phylogenesis, rapid molecular identification and the like of the cerasus schneideriana.

The invention discloses a novel coronavirus nucleic acid detection kit, which belongs to the technical field of biological detection. The present invention uses the real-time fluorescence quantitative cross primer isothermal amplification technology to detect the novel coronavirus, and designs two sets of primers suitable for this technology. The two sets of primers are respectively for the detection of the ORF1ab gene in the type coronavirus genome and the S gene in the type coronavirus genome. The novel coronavirus nucleic acid detection kit of the invention has strong specificity and high sensitivity, not only can qualitatively and quantitatively detect samples, but also can observe changes in data in real time, which is conducive to the comprehensive collection of sample data and facilitates further research. Therefore, the present invention is not only suitable for on-site rapid detection, but also for detection for the purpose of scientific research.

The invention discloses a DNA barcode, the nucleotide sequence of which is shown in SEQ ID NO.1. The DNA barcode disclosed the ITS sequence of Muscadine grape for the first time, which provided an effective molecular marker for the species identification of Muscadine grape. In addition, the DNA barcode also included some conserved sequences, which is universal when applied to the species identification of grapevine. The invention discloses a method for preparing the nucleotide sequence of the DNA barcode and primers, which can be used to obtain the sequence of the ITS region of muscadine grapes. The invention also discloses a molecular identification method of muscadine grapes, detection primers and a kit for identifying muscadine grapes, using the detection primers or kits to carry out PCR amplification to obtain the sequence of the grape ITS2 region, and then combining the amplified sequence with the muscadine The DNA barcode sequences of grapes are compared or applied to construct a phylogenetic tree of grapes to achieve accurate identification of grape varieties, and the identification process is simple, efficient and repeatable.

The invention relates to the field of molecular biology, particularly a primer set and application thereof in amplifying SIV / SHIV (simian immunodeficiency virus / simian human immunodeficiency virus) genome, and a kit. The primer set provided by the invention can be used for specific, stable and accurate amplification to obtain the target sequence. The agarose gel electrophoresis detection indicates that no non-specific strip is generated, and no trailing or dispersion is generated. Compared with the SIVmac239 whole genome sequence registered in Genbank after sequencing, the matching degree is 100%, and no base replacement or deletion (gap) is detected. The experiment indicates that the primer set has favorable sensitivity for the SIV / SHIV genome and can have favorable detection effects when the virus copy number is 100.5-1.0, and thus, the analysis judges that the sensitivity difference is related to virus sequence variation degree.

The present invention discloses a detection method of a turbot FF0922 microsatellite marker by utilizing specific primers, comprising the following steps: firstly extracting turbot genomic DNA; then utilizing a sequence containing a microsatellite in an enrichment library of the turbot microsatellite; designing specific primers at two ends of the sequence as a positive strand 5'-TGTGAGAAAACAGGCAATC-3', and a negative strand CAGTTTCTGTGATTTGGTGAT-3'; carrying out PCR amplification on the turbot genomic DNA; separating the obtained PCR product through modified polyacrylamide gel electrophoresis; analyzing strips generated on the product to obtain a turbot genetic polymorphism map. The method of the invention is suitable for the detection technologies of turbot population genetic marker, genealogy authentication, genetic linkage map construction and the like; the PCR amplification product presents better polymorphism in turbot population detection; and the method of the invention can rapidly acquire a genetic variation map of the turbot FF0922 microsatellite marker, and conveniently, rapidly, accurately and intuitively detect each individual genotype of the turbot.

The invention discloses a primer and method for detecting the mononucleotide polymorphism of a DQA1 gene related to goat immunity. The primer comprises an upstream primer and a downstream primer. When detection is carried out, goat genome DNA is taken as a template, PCR (polymerase chain reaction) is carried out by utilizing the upstream primer and the downstream primer to amplify a target fragment, a PCR product is treated by using a denaturing agent, the denatured amplified fragment is subjected to non-denatured polyacrylamide gel electrophoresis detection, glue dyeing is carried out by applying a silver dyeing method, and an allelic gene of a goat DQA1 gene exon 4 can be judged according to a polyacrylamide gel electrophoresis result. According to the nucleotide primer disclosed by the invention and an established detection system, different allelic genotypes of a group can be more accurately and faster amplified, and different mononucleotide mutant sites of the goat DQA1 gene exon 4 can be obtained through accurate analysis.

The invention discloses a detection method of a turbot FF0901 microsatellite marker by utilizing specific primers, comprising the following steps: firstly extracting turbot genomic DNA; then utilizing a sequence containing a microsatellite in an enrichment library of the turbot microsatellite; designing specific primers at two ends of the sequence as a positive strand 5'-TTACGCTTCGCATCGCTCAT-3', and a negative strand 5'-TGCCAGGCCACCAGACG-3'; carrying out PCR amplification on the turbot genomic DNA; separating the obtained PCR product through modified polyacrylamide gel electrophoresis; analyzing strips generated on the product to obtain a turbot genetic polymorphism map. The method of the invention is suitable for the detection technologies of turbot population genetic marker, genealogy authentication, genetic linkage map construction and the like; the PCR amplification product presents better polymorphism in turbot population detection; and the method of the invention can rapidly acquire a genetic variation map of the turbot FF0901 microsatellite marker, and conveniently, rapidly, accurately and intuitively detect each individual genotype of the turbot.

The invention discloses a viral hepatitis treatment drug interferon production technology which comprises the following steps: (1) acquisition of a target gene; (2) amplifying by PCR (polymerase chain reaction), to be more specific, in a microcentrifuge tube, using the 1 g / mL target gene obtained by the step (1) as template DNA, adding a right amount of a buffer solution, 4 kinds of dNTP (diethyl-nitrophenyl thiophosphate) mixtures, 0.5 2.5U / 50 muL DNA polymerase system, a pair of 0.1-0.5 mumol / L synthesized DNA primer to make the system pH 6.8-7.8; (3) gene recombination; (4) transformation; (5) separation and purification; (6) gene expression; (7) modification; and (8) freeze-drying, mass viral hepatitis treatment drug interferon can be produced in a short period of time, after modifying with polyethylene glycol, the half-life of the interferon is prolonged, hepatitis virus targeting property is improved, the antigenicity is weak, no human body immune response is caused, drug efficacy is good, production is fixed, stability is good, and the viral hepatitis treatment drug interferon production technology is suitable for popularization and application.

The invention discloses a method for quantitatively detecting endophytic bacteria of plant tissues through a non-culture method. The method comprises the following steps: adding bacterial DNA with different copy numbers into genome DNA extracted from sterile calluses, and carrying out fluorescent quantitative PCR (Polymerase Chain Reaction) by using a bacterial 16S rRNA gene specific primer pair to obtain a standard curve of the plant tissue bacterial content and a fluorescent quantitative PCR reaction Ct value; carrying out surface sterilization on plant tissues to be detected, and extracting total DNA; and taking the extracted DNA as a template, carrying out fluorescent quantitative PCR (Polymerase Chain Reaction) by adopting the bacterial 16S rRNA gene specific primer pair, and substituting the obtained Ct value into the standard curve to obtain the content of the endophytic bacteria in the plant tissue to be detected. The method is easy and convenient to operate, time-saving, efficient, capable of covering all endophytic bacteria of plant tissues and suitable for various plants, and the bacterial content of the plant tissues to be detected can be directly obtained by substituting the qPCR reaction Ct value of a sample to be detected into the standard curve in related research.

The invention discloses a primer set for rapidly detecting human DYS385 gene. Wherein, the primer set is composed of the following primers: primer F1 as shown in SEQ ID No.1; primer B1 as shown in SEQ ID No.2; primer F2-1 as shown in SEQ ID No.3; primer B2-1 As shown in SEQ ID No.4; Primer F2-2 as shown in SEQ ID No.5; Primer B2-2 as shown in SEQ ID No.6; Primer LOOP-F as shown in SEQ ID No.7; Primer LOOP -B is shown in SEQ ID No.8. The detection time required by the present invention is short, and the corresponding primers and kit can quickly detect the human DYS385 gene within 30 minutes to determine the sex of the human sample.

The invention discloses primers and a kit for detecting the macrolide drug-resistance genes of bacteria, and belongs to the technical field of microbiological detection. The sequences of the primers for detecting the macrolide drug-resistance genes of bacteria provided by the invention are shown in SEQ ID NO: 1-6, and the three primer pairs are capable of specifically amplifying the three macrolide drug-resistance genes of ermA, ermB and ermC of multiple bacteria. The invention further discloses a kit for detecting the macrolide drug-resistance genes of bacteria, wherein the kit contains an EvaGreen fluorescent dye, and is capable of rapidly and accurately detecting the macrolide antibiotic-related drug-resistance genes of bacteria. The primers and the kit disclosed by the invention have the advantages of simplicity and convenience in operation, high specificity, high sensibility, low cost, high flux and the like, and can be used for rapidly detecting macrolide antibiotic-related drug-resistance genes of clinic pathological bacteria and providing a reference for clinic treatment.

The invention relates to a scophthalmus maximus T170G single nucleotide polymorphic marking detection method. The method comprises the following steps: extracting scophthalmus maximus genome DNA and diluting for later use; analyzing the sequences of EST, screening the sequence containing a candidate SNP locus, designing a specific primer at the two ends and designing a non-marking probe with the closed 3' end in front of and behind the locus (comprising the locus); performing asymmetric PCR amplification on the scophthalmus maximus group genome DNA by using the primer; hybridizing the amplification product with the non-marking probe with the closed 3' end; and placing the hybrid product on a LightScanner, and detecting and analyzing the melting curve to obtain the genetic polymorphism map of the scophthalmus maximus. The method is applicable to detection technologies of scophthalmus maximus genetic marking, genealogy authentication, genetic linkage map construction and the like. According to the method which is convenient, quick and accurate, the scophthalmus maximus T170GSNP marked genetic variation map can be obtained quickly; and the genotype of each individual of the scophthalmus maximus can be detected intuitively.

The invention discloses a poppy species specificity genetic marker detecting system. A primer pair set for identifying poppy is provided and is composed of a primer pair P12 (sequence 1 and 2), a primer pair P13 (sequence 3 and 4), a primer pair P5 (sequence 5 and 6) and a primer pair P14 (sequence 7 and 8). Four STR loci of poppy are amplified at the same time, and the amplification is more accurate and rapid compared with amplification of a single locus; the system has higher species specificity when identifying poppy and affinis plants, and an effective method is provided for further studying poppy STR loci and inspecting and identifying the poppy species in a case; in an optimized poppy STR composite amplification system, amplification of all loci can be balanced, sensitivity is high, stability is high, and frequently-seen corrosion check materials in a drug-related case can be easily inspected; accuracy is high, parting of four loci of each of poppy samples from different producing areas can be achieved through detection and data statistic analysis.