Tegillarca granosa mitochondria COI gene amplification primers

A mitochondrial and COINR technology, applied in the direction of DNA preparation, recombinant DNA technology, etc., can solve the problems of time-consuming, labor-intensive, high cost, unsuitable for cockle population genetics research, etc., and achieve the effect of high amplification efficiency

Inactive Publication Date: 2012-11-07
OCEAN UNIV OF CHINA
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Problems solved by technology

Although this method can obtain more accurate sequencing results, it is time-consuming, laborious, and expensive, and is not suitable for large-scale cockle population genetics research

Method used

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  • Tegillarca granosa mitochondria COI gene amplification primers

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Embodiment Construction

[0008] When implementing the screening method of the clam mitochondrial COI gene amplification primer of the present invention, divide the following five steps:

[0009] a. Log in to the National Center for Biotechnology Information (NCBI, http: / / www.ncbi.nlm.nih.gov / ), and download the existing 39 cockle mitochondrial COI sequences (Genbank accession numbers are: EF583524-583540 and FJ411459 -411480).

[0010] b. Use the sequence processing software BioEdit (http: / / www.mbio.ncsu.edu / BioEdit / bioedit.html) to perform multiple alignment analysis on the above sequence, and determine the more conserved regions at the front and back ends.

[0011] c. After comparison, use the primer design software Primer Premier 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, and design primers Try to avoid the occurrence of hairpin structure and primer dimer. The designed multiple pairs...

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Abstract

The invention screens a pair of primers for amplification of a tegillarca granosa mitochondria COI sequence. The method comprises the following steps of: downloading the tegillarca granosa mitochondria COI sequence existing in the National Center of Biotechnology Information (NCBI); comparing and analyzing the tegillarca granosa mitochondria COI sequence by using BioEdit software; after determining a conserved domain, designing a plurality of pairs of primers by using primer design software Primer Premier 5; performing PCR reaction between the DNA of individual tegillarca granosa and the plurality of pairs of synthesized primers in a reaction system respectively; and finally, performing agarose gel electrophoresis detection to screen out a pair of primers COINF and COINR. Tests prove that the pair of primers has the advantages of single stripe and over 90-percent amplification efficiency, can meet the requirements of study on the genetic diversity of the tegillarca granosa, and plays an important role in the protection and utilization of tegillarca granosa resources, diversity estimation, genetic breeding and the like.

Description

technical field [0001] The invention relates to a screening method for amplifying primers of cockle mitochondria cytochrome oxidase subunit I (COI) gene. Background technique [0002] Mud cockle (Tegillarca granosa), commonly known as blood cockle, belongs to Mollusca, Lanellibranchia, Taxodonta, Arcidae, and Tegillarca Iredale. The bivalve molluscs in the intertidal muddy deposit areas are distributed in all coastal provinces and cities of the country. [0003] Mud cockles are one of the four major cultured shellfish. The meat is delicious and can be eaten fresh or made into dried products. The cockle meat is rich in protein and vitamins. As a popular seafood product, cockles are artificially cultured in Hebei, Shandong, and southern provinces and cities in my country, and the output is quite abundant. There have been many reports on artificial breeding, reproduction and growth, physiology and biochemistry of cockles, but few studies on genetic diversity based on molecula...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
Inventor 倪刚李琪孔令锋
Owner OCEAN UNIV OF CHINA
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