Universal primers for cephalopod mitochondrial COI genes and design and amplification method thereof

A universal primer and cephalopod technology, which is applied in biochemical equipment and methods, microbial measurement/testing, DNA/RNA fragments, etc., can solve the problems of inability to identify cephalopod mitochondrial COI gene, time-consuming, inconvenient and efficient, etc. , achieve the effect of cost saving and simple operation

Inactive Publication Date: 2018-04-20
ZHEJIANG OCEAN UNIV
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Problems solved by technology

[0004] On September 29, 2017, the Chinese Patent Office disclosed the invention authorization of a spider mitochondrial genome full-sequence amplification primer and amplification method, the authorization publication number: CN104531688B. Long PCR (L-PCR) amplification and sequencing identify spiders, which is similar to the present invention, but it can only identify spider mitochondrial DNA, but cannot identify cephalopod mitochondrial COI genes, and it is necessary to design multiple pairs of primer pairs Amplification of the entire genome sequence takes a long time and is not convenient and efficient

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  • Universal primers for cephalopod mitochondrial COI genes and design and amplification method thereof

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Embodiment

[0022] (1) Collection, identification and preservation of cephalopod samples

[0023] The samples of cephalopods used in the examples were all collected from the natural environment in the wild, and all the collected samples were brought back to the laboratory for information registration, and all were identified with a stereo microscope to determine the species. The identified cephalopod samples were soaked in 100% pure alcohol and stored in a refrigerator at 4°C for later use. The species of cephalopods collected include: Pacific Northwest squid, fluorescent cuttlefish, Argentine squid, king squid, curly squid, deep-sea squid, half monkhead squid, lychee squid, opalescent squid, mansoni squid and sorrel octopus.

[0024] (2) Extraction of cephalopod mitochondrial COI gene

[0025] The DNA extraction kit method was used to extract the mitochondrial COI gene of the cephalopod samples to be tested. In order to avoid the contamination of intestinal contents, the head and neck ...

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Abstract

The invention relates to universal primers capable of highly efficiently and quickly amplifying cephalopod mitochondrial COI genes of various cephalopods, and a design and amplification method thereof. By designing the universal primers of the cephalopod mitochondrial COI genes, various cephalopod COI genes can be amplified at a time in batches, specific primers are not needed to be designed for acertain single species, the time cost is saved, the work efficiency is greatly improved, and the method has high-efficiency and fast characteristics and provides important molecular means and data accumulation for species identification, systematic evolution, germplasm resource conservation and sustainable development and utilization of the cephalopods.

Description

technical field [0001] The invention relates to the field of cephalopod mitochondrial genome research, in particular to a general primer capable of efficiently and quickly amplifying cephalopod mitochondrial COI genes of various cephalopods and a method for designing and amplifying them. Background technique [0002] Cephalopods are an important group of molluscs. They are widely distributed in the three oceans of the world and the Antarctic waters. They not only have high economic value, but also occupy an important position in marine ecology. There are 2 subclasses, 11 orders, 50 families, and 154 genera and about 718 species of cephalopods. Since the 1970s, due to the general decline of traditional economic species resources in the world, the cephalopod resources at the low end of the food chain have attracted attention from countries all over the world. regional attention. According to the statistics of FAO, since 2000, the output of cephalopods in the world is about 31...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2600/156C12Q2563/185
Inventor 龚理吕振明刘立芹陈永久杜珣姜辉
Owner ZHEJIANG OCEAN UNIV
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