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A primer for amplification of mitochondrial cox2 gene of common northern clams and its screening method

A technology for amplification primers and screening methods, which is applied in the screening field of amplification primers for the mitochondrial cox2 gene of razor clams common in northern China, can solve the problems of no reports on razor clams, and achieve the effect of single band and high amplification efficiency

Inactive Publication Date: 2018-03-20
山东省海洋资源与环境研究院
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AI Technical Summary

Problems solved by technology

At present, the mtDNA of razor clam, razor clam, and razor clam have been determined, but the mtDNA sequence of razor clam has not been reported yet.

Method used

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Embodiment Construction

[0007] When implementing the screening method of northern common razor clam mitochondria cox2 gene amplification primer of the present invention, divide following five steps:

[0008] a. Log in to the National Center for Biotechnology Information (NCBI, http: / / www.ncbi.nlm.nih.gov / ), and download the cox2 sequences of the existing complete mitochondrial sequences of razor clams, razor clams, and razor clams (Genbank login No.: HQ703012, JN786377 and JN398366).

[0009] b. Use the sequence processing software BioEdit (http: / / www.mbio.ncsu.edu / BioEdit / bioedit.html) to conduct multiple alignment analysis on the above sequence, and determine the more conserved regions at the front and back ends.

[0010] c. After comparison, use the primer design software PrimerPremier 5 (http: / / www.premierbiosoft.com / primerdesign / index.html) to design primers in the relatively conserved regions of 100 base pairs at the front and rear ends. When designing primers Try to avoid the appearance of ha...

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Abstract

The present invention relates to a common northern razor clam mitochondrial cox2 gene amplification primer and its screening method. It is divided into five steps: a. Log in to the National Biotechnology Information Center of the United States, and download the cox2 in the complete mitochondrial sequence of razor clam, long razor clam, and constricted razor clam Sequence; b. Compare and analyze the downloaded sequence with software to determine the conserved regions at both ends of the sequence; c. Design multiple pairs of primers in the determined conserved region with primer design software and send them to Shanghai Sangon Biotechnology Co., Ltd. for synthesis; d. Extract DNA from the individuals of razor clam, razor clam, razor clam and razor clam, and then carry out PCR reactions with multiple pairs of primers synthesized in the reaction system to detect the amplification of the primers; e, carry out agarose gel electrophoresis detection, and screen A pair of primers cox2CF and cox2CR with single band and high amplification efficiency were obtained. The invention screens out a pair of primers with the characteristics of single band and high amplification efficiency, helps to quickly obtain the full length of mitochondrial genome, and satisfies the needs of genetic diversity and phylogenetic research of razor clams.

Description

technical field [0001] The present invention relates to a shellfish mitochondrial cox2 [cytochrome c Oxidase subunit II (cytochrome c oxidase subunit II)] gene amplification primer screening method, particularly relates to a screening method for the northern common razor clam mitochondrial cox2 gene amplification primer. Background technique [0002] Razor clams belong to the phylum Mollusca, class Bivalvia, subclass Heterodonta, and order Veneroida. They live in the intertidal zone to a water depth of about 400m. The meat is delicious and has High food value. Especially large razor clams, long razor clams, small knife razor clams and hanging razor clams are widely distributed in my country and are important economic shellfish. [0003] In recent years, due to overharvesting, deteriorating marine environment and shrinking habitat, the resources of razor clams have been declining year by year. It is urgent to speed up the assessment of its diversity to lay a foundation for...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/10
Inventor 冯艳微刘相全韦秀梅姜绪王卫军
Owner 山东省海洋资源与环境研究院
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