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Amplification primer for grouper catostomidae mitochondrial genome complete sequence and design method thereof

A mitochondrial genome and amplification primer technology, applied in the field of grouper, can solve the problems of high difficulty, cumbersome procedures, slow progress of determination work, etc., and achieves the effects of good versatility and high amplification efficiency

Inactive Publication Date: 2010-04-28
XIAMEN UNIV
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Problems solved by technology

However, due to the lack of primers suitable for the amplification and sequencing of the complete mitochondrial genome sequence, the determination of the complete mitochondrial genome sequence of the grouper is progressing slowly
Meyer (4, MeyerA (1993) Evolution of mitochondrial DNA in fishes.In: Biochemistry and Molecular Biology of Fishes (eds.Hochanchka PW, MommsenTP), vol.2, 1-38, Elsevier, NewYork) once announced in 1993 that fish General amplification primers for mitochondrial genomes, but grouper mitochondrial genome sequences are relatively specialized, and it is difficult to effectively amplify them with general amplification primers
So far, only Zhu et al. (5, Zhu ZY, Yue GH (2008) The complete mitochondrial genome of red grouper Plectropomus leopardus and its applications in identification of grouper species. Aquaculture, 276, 44-49) successfully utilized Meyer's universal amplification primers Amplify the complete mitochondrial genome sequence of a grouper fish, and these primers have high requirements on the quality of DNA templates, long-chain PCR amplification and step-by-step sequencing, which are difficult and cumbersome

Method used

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Embodiment Construction

[0059] The primers for amplification of the complete mitochondrial genome sequence of grouper subfamily fishes according to the present invention are composed of 16 pairs of single-stranded oligonucleotides, and PCR amplification is performed on 32 kinds of grouper, and the correct target fragment can be obtained. In order to quickly obtain the full sequence of its mitochondrial genome. Therefore, it provides a powerful tool for the species identification, geographic population identification and germplasm resource evaluation of groupers in my country, and makes rapid, accurate and efficient identification of grouper species, geographic populations and Germplasm resource assessment becomes a reality. The 32 species of groupers involved cover a total of 6 genera of species, which are:

[0060] 1 Gem Grouper, 2 Bruce's Grouper, 3 Double-Spine Grouper, 4 Long-Spine Grouper, 5 Red Spot Grouper, 6 Hawksbill Grouper, 7 Electric Stripe Grouper, 8 Salmon Spot Grouper, 9 black-edged g...

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Abstract

The invention discloses an amplification primer for a grouper catostomidae mitochondrial genome complete sequence and a design method thereof, relates to groupers, and particularly relates to 16 pairs of amplification primers mainly used for specifically amplifying most grouper catostomidae mitochondrial genome complete sequences and a design method thereof. The invention provides an amplification primer for a grouper catostomidae mitochondrial genome complete sequence, a design method thereof and application thereof. The amplification primer for the grouper catostomidae mitochondrial genome complete sequence consists of 16 pairs of single-chain oligonucleotides, wherein each pair consists of one light-chain primer and one heavy-chain primer. The amplification primer for the grouper catostomidae mitochondrial genome complete sequence can be used in the aspects of variety authentication, geographical population authentication, germplasm resource evaluation and the like.

Description

technical field [0001] The invention relates to grouper, in particular to 16 pairs of amplification primers mainly used for specifically amplifying the complete mitochondrial genome sequence of most grouper subfamily fishes and a design method thereof. Background technique [0002] The mitochondrial genome consists of 37 coding genes and a major non-coding region (control region), and the evolution rates of different sequence regions are different. Because animal mitochondrial DNA (mtDNA) has the characteristics of simple structure, strict maternal inheritance, almost no recombination, fast evolution rate and differences in evolution rate in different regions, it is widely used in species identification, phylogenetic evolution and population genetics research. Very broad. [0003] At present, it is a very convenient and accurate method to identify fish with similar appearance through sequence differences. Among them, the study of mitochondrial sequence differences, such as ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 丁少雄曾华嵩庄轩
Owner XIAMEN UNIV
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