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Molecular specific marker primer for identifying Chinese sturgeon, and method

A technology for labeling primers and Chinese sturgeon, applied in DNA/RNA fragments, recombinant DNA technology, etc., can solve the problem of lack of data, and achieve the effect of simple method and high sensitivity

Inactive Publication Date: 2018-03-23
ZHENJIANG HUADA DETECTION CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the endemism of many fishes in our country, there is not enough relevant data

Method used

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  • Molecular specific marker primer for identifying Chinese sturgeon, and method
  • Molecular specific marker primer for identifying Chinese sturgeon, and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] (1) Extraction of Chinese sturgeon genome DNA to be tested

[0034] The Chinese sturgeon tissue samples to be tested were cut up and extracted using a genomic DNA extraction kit. The DNA obtained was detected by electrophoresis using 1.5% agarose gel, and the DNA concentration was detected by an ultraviolet spectrophotometer, and stored in a -20°C refrigerator for later use.

[0035] (2) Design and synthesize specific PCR amplification primers

[0036] Design and synthesis of specific primers based on Chinese sturgeon, the upstream primer XYF: 5'-GCCATTCGTAATCGCCGGAGC-3' and the downstream primer XYR: 5'-AGCATTAAGATAAATCCGAAG-3', were designed by technicians and synthesized by the primer design company.

[0037] (3) PCR amplification of molecular specific marker primers (XYF / XYR)

[0038] 25 μL of PCR reaction system: 2.5 μL of 10×PCR Buffer, 0.5 μL of dNTPS, 0.2 μL of Taq (5U / μl), MgCl 2 1.5 μL, XYF primer (10 μM) 1 μL, XYR primer (10 μM) 1 μL, DNA template (about 1...

Embodiment 2

[0044] The method in Example 1 was used to detect three species of Chinese sturgeon, Russian sturgeon, and crucian carp. The sampler recorded and numbered the species according to the specific information, and carried out the experiment when the experimenter did not know the species name. The results showed that only one kind of fish could be amplified. A specific DNA band with a molecular weight of about 150bp was obtained, but the other two fish samples could not be amplified, and the test results were completely consistent with the information recorded by the sampler. It can be shown that the nucleic acid molecule primers of the present invention have extremely high specificity, and therefore can be used for rapid identification of Chinese sturgeon.

Embodiment 3

[0046] Two groups of muscle tissue samples were selected from the sturgeon tissue samples and preserved in 95% alcohol. The selected muscle tissues belong to two groups of sturgeons of different species, one of which is Chinese sturgeon (5 samples), and the other group is other species (5 samples), and the sampler shall record and number the species according to the specific information , the experimental operator conducted the experiment without knowing the species of the two groups of samples. The same experimental method as in Example 1 was used to carry out the experiment, and the PCR product sequencing results showed that 5 samples of one group could amplify a specific DNA band of about 150bp, which was the tissue sample of Chinese sturgeon, and 5 samples of the other group It is a non-Chinese sturgeon tissue sample, and the experimental results are completely consistent with the information recorded by the sampler.

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Abstract

The invention discloses a molecular specific marker primer for identifying Chinese sturgeon, and a method. In the molecular specific marker primer for identifying the Chinese sturgeon, the upstream primer XYF is shown in SEQ ID NO.1; and the downstream primer XYR is shown in SEQ ID NO.2. The molecular specific marker primer XYF / XYR performs rapid molecular identification on the Chinese sturgeon bythe conventional PCR method, the method is simple, detection is conducted by a PCR technology, the time is short, detection can be completed in half a day, and the molecular specific marker primer and the method are effective assistance for morphological character discrimination on the Chinese sturgeon.

Description

technical field [0001] The invention relates to a molecular specific marker primer for identifying Chinese sturgeon, and a method for quickly identifying Chinese sturgeon specimens and individuals by using the specific primer. Background technique [0002] Chinese sturgeon (scientific name: Acipenser sinensis) is a large anadromous fish with a maximum weight of 550 kg. It usually lives in the eastern coast of my country. After sexual maturity, it migrates into rivers for reproduction. The spawning grounds are mainly distributed in the Yangtze River. In addition, a small number of Chinese sturgeons were also found to lay eggs in the Pearl River. [0003] DNA barcoding technology (DNA barcoding) refers to a new molecular identification technology that uses a standard, short DNA fragment in the genome to identify species, which can quickly and accurately identify species. A common molecular marker in animals is the partial sequence of the mitochondrial cytochrome b gene (Cytb) ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
Inventor 张继敏耿小燕崔海月游欣欣王敏朱志飞
Owner ZHENJIANG HUADA DETECTION CO LTD
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