37results about How to "Accurate amplification" patented technology

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 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 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.