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Spider mitochondria ND4 gene complete sequence amplification primer and identification method thereof

A technique for amplifying primers and mitochondria, which is applied in biochemical equipment and methods, microbiological determination/testing, recombinant DNA technology, etc., to achieve the effects of low equipment requirements, overcoming difficulties, and shortening the identification cycle

Inactive Publication Date: 2015-03-25
CHINA JILIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

However, amplified mitochondria ND4 Universal primers for gene sequences are seldom reported, especially for a wide variety of spiders with important utilization value

Method used

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  • Spider mitochondria ND4 gene complete sequence amplification primer and identification method thereof

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Embodiment 1

[0022] (1) Collection, identification and preservation of spider samples

[0023] The species of spiders used in the examples were all collected from the farmland of Yuyao City, Zhejiang Province by trap method. The collected specimens were brought back to the laboratory for information registration, and were identified with a stereo microscope to determine the species. The spider specimens used for molecular biology experiments were soaked in 100% alcohol and stored in a 4°C refrigerator for later use. The species of spiders collected include: Pseudomonas tarantula, Triflower spider, Golden spider, Cone-bellied octopus, Ornate octopus, Brown new orb spider, Maiwa spider, black fly tiger, big-bellied garden spider, cat jumping spider.

[0024] Extraction of Total DNA from Spider Genome

[0025] 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 extract...

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Abstract

The invention discloses a spider mitochondria ND4 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 ND4 gene complete sequence amplification primer, performing PCR amplification and sequencing, and rapidly identifying the species of spiders based on an ND4 base sequence. The complete sequence of the multiple mitochondria ND4 genes of the spiders can be rapidly, efficiently and specifically amplified, a powerful tool can be provided for rapid identification of the species of spiders and population diversity analysis, and the method can be applied to analysis and research on the revolutions of different taxonomic category systems of the spiders.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a primer for amplifying the full sequence of a spider mitochondrial NADH reductase complex subunit 4 (NADH dehydrogenase subunits 4; ND4) gene and an identification method. Background technique [0002] Spiders are important predatory natural enemies of pests and play an important role in the process of integrated pest management. Attention should be paid to protecting the natural enemy species pool and promoting 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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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 俞晓平王正亮李超
Owner CHINA JILIANG UNIV
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