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Amplification primer of psammobiidae shellfish mitochondria COI (Cytochrome Oxidase I) gene

A technique for amplifying primers and mitochondria, applied in DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of insufficient research on genetic diversity and obstacles to the development, utilization and protection of Ziyun clam germplasm resources

Inactive Publication Date: 2012-11-14
OCEAN UNIV OF CHINA
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Problems solved by technology

[0003] There have been reports on the breeding and biological characteristics of Ziyun clam, but there are few studies on genetic diversity based on molecular markers. This situation greatly hinders the development, utilization and utilization of Ziyun clam germplasm resources. Protect

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  • Amplification primer of psammobiidae shellfish mitochondria COI (Cytochrome Oxidase I) gene

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Embodiment Construction

[0007] The screening method of the amplification primer of the Mitochondriaceae Mitochondrial COI gene of the present invention adopts three steps: a, utilize the COI universal amplification primer (LCO1490F and HCO2198R, Folmer 1994) to amplify the COI sequence of a part of the Mitochondriaceae species b, from the part of the Clamidae COI sequence obtained above, find out the conserved sequence fragments by comparison, and synthesize multiple sets of amplification primer sequences; c, screen out the amplification from the multiple sets of synthetic amplification primer sequences A pair of primers with the best effect on the mitochondrial COI gene of the family Clamidae.

[0008] The present invention will be further described below in conjunction with embodiment:

[0009] 1. Sample collection: From 2002 to 2010, a total of 200 individuals of 10 species belonging to 4 genera and 10 species of Clamidae were collected from the north and south coasts of China.

[0010] 2. DNA ex...

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Abstract

The invention relates to an amplification primer of a psammobiidae shellfish mitochondria COI (Cytochrome Oxidase I) gene. 64 psammobiidae sequences obtained by amplification of universal primers are subjected to comparative analysis by using CLUSTALW software; a plurality of pairs of primers are designed by using primer design software Primer Premier 5 in a determined conserved region; PCR (Polymerase Chain Reaction) reaction experiments are repeatedly performed by using 200 psammobiidae individuals under a 10 mu system; and finally an efficient amplification primer of a psammobiidae shellfish mitochondria COI gene sequence is sifted through detection of agarose gel electrophoresis. The amplification primer is characterized in that COIZGF is 5'-ACAAATCATTAAGGATATTGG-3'; and COIZGR is 5'-GTCCGAAGAATCAAAACAAATG-3'. With the adoption of the primer of the psammobiidae shellfish mitochondria COI, provided by the invention, target segments of different species of psammobiidae shellfishes can be effectively amplified, the amplification efficiency is more than 90%, the primer can satisfy the demands of genetic diversity research on psammobiidae, and plays an important role in the aspects of protection and utilization of psammobiidae resources, diversity assessment, genetic breeding and the like.

Description

technical field [0001] The invention relates to a primer for amplifying the mitochondrial COI [the cytochrome c oxidase I] gene of the clam family. Background technique [0002] Corbiculacea belongs to Bivalvia, Heterodonta, Veneroida, all marine products, widely distributed in coastal China, Japan, Philippines and Indian Ocean. There are many species of shellfish in this family, and about 25 species are known in China, usually buried in the intertidal zone to shallow sea fine sand or muddy sea bottom. Most of the shellfish of Ziyun clam family have delicious meat, high nutritional value and important economic value. For example, Chinese purple clams, double-lined purple clams, Zicai blood clams and pointed purple clams are all rare and precious sea treasures. However, due to the high degree of plasticity of the external shell morphology and the high similarity of the mantle sinus morphology of the species of the family Clamidae, it is extremely difficult to use morphologi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11
Inventor 于贞贞李琪孔令锋
Owner OCEAN UNIV OF CHINA
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