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A kind of amplification primer and screening method of mitochondrial cox3 gene of common northern razor clams

An amplification primer, mitochondrial technology, applied in DNA preparation, DNA/RNA fragments, recombinant DNA technology, etc., can solve problems such as hindering the acquisition of full length, and achieve the effect of single band and high amplification efficiency

Inactive Publication Date: 2018-06-15
山东省海洋资源与环境研究院
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Problems solved by technology

In the process of measuring the mtDNA of razor clam by walking method, a pair of Long-PCR primers designed using short fragments of cox1 gene and 16S rRNA gene failed to amplify successfully, which hindered the acquisition of full-length

Method used

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Embodiment Construction

[0007] When implementing the amplification primers of the mitochondrial cox3 gene of common razor clams in the north of the present invention, the following five steps are performed:

[0008] 1. Download the sequence: log in to the National Center for Biotechnology Information (NCBI, http: / / www.ncbi.nlm.nih.gov / ) to download the existing cox3 sequence in the mitochondrial sequence of the razor clam, razor clam, and razor clam (Genbank accession numbers are: HQ703012, JN786377 and JN398366).

[0009] 2. Sequence alignment: Use the sequence processing software BioEdit (http: / / www.mbio.ncsu.edu / BioEdit / bioedit.html) to perform multiple alignment analysis on the above sequences to determine the more conservative regions in the front and back ends.

[0010] 3. Primer design and synthesis: After comparison, use the primer design software Primer Premier 5 (http: / / www.premierbiosoft.com / primerdesign / index.html) in the more conservative region of 100 base pairs in the front and back ends. Wh...

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Abstract

The invention relates to an amplification primer and a screening method for a common northern razor clam mitochondrial cox3 gene. The amplification primer cox3CF is 5' TATCATTTAGTHGATATAAGACC 3'; the amplification primer cox3CR is 5' CCATGAAADCCAGTTATYACAA 3'; the preparation method of the amplification primer comprises Download sequence, sequence comparison, primer design and synthesis, DNA extraction, primer amplification five steps; the present invention screens out a pair of primers cox3CF and cox3CR for amplifying the cox3 sequence of the common northern razor clam mitochondria, and determines its reaction System and PCR amplification conditions, experiments have proved that the primer has the characteristics of a single band and high amplification efficiency, and can meet the needs of the research on the mitochondrial genome of razor clams and the subsequent research on genetic diversity and phylogenetics.

Description

Technical field [0001] The present invention relates to a shellfish mitochondrial cox3 [cytochrome c Oxidase subunit III (cytochrome c oxidase subunit III)] gene amplification primers, especially related to a common razor clam mitochondrial cox3 gene amplification primer and its screening method. Background technique [0002] Razor clams belong to Mollusca, Bivalvia, Heterodonta, Veneroida. They live in intertidal waters up to hundreds of meters deep. They are common economic shellfish. class. In particular, razor clams, long razor clams, razor clams and constrictor clams are widely distributed in our country, and they are one of the favorite marine shellfish of the coastal people of our country, and occupy an important position in the edible shellfish. [0003] In recent years, due to over-harvesting, deteriorating marine environment and shrinking habitats, the resources of razor clams have been declining year by year. It is urgent to accelerate the assessment of its diversity an...

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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