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Spider mitochondria COIII gene complete sequence amplification primer and identification method thereof

A technology of COIII-F and amplification primers, which is applied in biochemical equipment and methods, measurement/inspection of microorganisms, DNA/RNA fragments, etc., can solve the problems that have not been reported, and the application of general primers hindering the amplification efficiency, etc. The effect of low equipment requirements, shortening the appraisal cycle, and overcoming difficulties

Active Publication Date: 2015-03-25
CHINA JILIANG UNIV
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Problems solved by technology

However, the current mitochondrial COIII Universal amplification primers for genes have not been reported
The lack of universal primers with high amplification efficiency severely hampers COIII Application of genes as molecular markers, thus engineered to specifically amplify spider mitochondria COIII The universal primer of the gene is of great significance to the rapid identification of spider species and the investigation and analysis of species diversity by molecular methods, and it also lays the foundation for the determination of the whole genome sequence and phylogenetic analysis of spider mitochondria

Method used

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  • Spider mitochondria COIII gene complete sequence amplification primer and identification method thereof

Examples

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Embodiment 1

[0021] (1) Collection, identification and preservation of spider samples

[0022] The species of spiders used in the examples were all collected from the farmlands of Yuyao City or Tongxiang County, Zhejiang Province by trap method. The collected specimens were brought back to the laboratory for information registration, and all were identified with a stereo microscope to determine the species. The spider specimens were soaked in 100% alcohol and stored in a 4°C refrigerator for later use. The species of spiders collected include: Pseudomonas tarantula, Triflora, Golden spider, Cone-bellied octopus, Ornate octopus, Brown new orb spider, Maiwa spider, black fly tiger, big-bellied garden spider, cat jumping spider.

[0023] Extraction of Total DNA from Spider Genome

[0024] DNA extraction kits were used to extract the total genomic DNA of different species of spiders. To avoid contamination, the head and abdomen were discarded, and DNA was extracted only from the legs....

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Abstract

The invention discloses a spider mitochondria COIII gene complete sequence amplification primer and a method for rapidly identifying species of spiders by utilizing complete sequence sequencing information. The method comprises the following steps: extracting total DNA of spider genomes, designing the COIII gene complete sequence amplification primer, performing PCR amplification, sequencing and rapidly identifying the species of spiders based on the gene complete sequence. With the adoption of the universal amplification primer disclosed by the invention, the complete sequence of the multiple mitochondria COIII genes of spiders can be rapidly and efficiently amplified, a powerful tool is provided for researches such as rapid identification of the species of spiders, population diversity analysis and genetic resource conservation and utilization, and a foundation is laid for spider mitochondria whole genome sequencing and analysis and research on different taxonomic category system evolutions.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a primer for amplifying the complete sequence of a spider mitochondrial cytochrome C oxidase 3 (cytochrome oxidase subunits 3; COIII) gene and a method for identifying spider species. Background technique [0002] Spiders are an important predatory natural enemy of pests and play an important role and status in the process of integrated pest management. Pay attention to the protection of the seed bank and promote the reconstruction of the natural enemy community, which can effectively control the occurrence and damage of many agricultural pests, such as planthoppers, leafhoppers, stem borers, and cotton aphids. Rapid and accurate species identification is the necessary premise and basis for the research on the protection and utilization of spiders. However, the traditional classification and identification methods based on appearance and anatomical features are not only...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 王正亮俞晓平李超
Owner CHINA JILIANG UNIV
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