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Northeast wood frog and molecular identification method for egg oil thereof

A molecular and analytical system technology, applied in the field of molecular identification of Northeast Rana species and its product egg oil, to achieve reliable detection results, strong specificity, and advanced and reasonable implementation steps

Inactive Publication Date: 2009-05-20
SHENYANG NORMAL UNIV
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  • Abstract
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Problems solved by technology

[0003] Aiming at the problem that it is difficult to identify the Northeast Rana and Rana egg oil in terms of traits, microscopic characteristics, physical and chemical properties, etc. in the production and management practice, the present invention uses molecular biology technology to solve the problem of Rana Rana egg oil and its medicinal source animal Northeast China. The molecular identification of wood frog provides a method based on PCR technology to screen specific primers for amplification, so as to achieve molecular identification of Northeast wood frog and its egg oil. The specific implementation steps are:

Method used

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  • Northeast wood frog and molecular identification method for egg oil thereof

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Embodiment

[0050] The authenticity of the Northeast forest frog and egg oil purchased from Changbai Mountain in Jilin were used as identification objects.

[0051] Three samples of Rana and egg oil were taken respectively. The specific operation steps are as follows:

[0052] (1) Genomic DNA extraction:

[0053] ①Use an electronic balance (FA1604N, Shanghai Precision Scientific Instrument Factory) to weigh 200 mg of sample tissue (the muscle tissue of the Northeast Rana sample), wash it with 70% alcohol, cut it into pieces with sterilized scissors, put it into a 1.5ml EP tube, and naturally Air dry for 30-60 minutes to evaporate the alcohol;

[0054] ②Add 600 μl of 1×SET solution, 20 μl of proteinase K (10 mg / ml), and 25 μl of SDS to the EP tube, and use an air bath shaker (CHA-S, Shenzhen Tiannanhaibei Co., Ltd.) at 55°C for 2 hours;

[0055] ③ Add 500 μl Tris saturated phenol and 200 μl TE to the EP tube and invert up and down for 10 minutes;

[0056] ④Use a high-speed centrifuge (...

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Abstract

A molecular identification method for the wood frog in the northeast of China and the ovum oil thereof aims at solving the identification of the wood frog in the northeast of China and the product ovum oil thereof, which is mainly based on the property characteristics, microscopic characteristics, physical and chemical properties, and the like at present; as the bases are lack of clear shape identification characteristics and special chemical compositions, the identification for the ovum oil in practice is still very difficult and the invention is hereby designed. The identification method takes PCR technology as the basis and screens specific primers for amplification so as to achieve the molecular identification of the wood frog in the northeast of China and the ovum oil thereof, comprising the following steps: (1) extraction of genomic DNA; (2) genomic DNA detection and concentration determination; (3) PCR amplification; and (4) PCR amplification product identification. The invention is characterized in that the invention starts from the basic theory of molecular biology, uses modern molecule marker technology for conducting molecular identification of the wood frog in the northeast of China and the ovum oil thereof, has the advantages of scientific and credible theoretical basis, advanced and reasonable technical means and achieving steps, reliable detection result, strong specificity of screened primers as well as high stability of agarose gel electrophoresis characteristic bands, and can obtain reliable and useful effects in practical application.

Description

Technical field: [0001] The invention relates to a biological molecular identification method, which is mainly aimed at the molecular identification method of the Northeast Rana species and its product egg oil. Background technique: [0002] Northeast Rana has important economic significance and medical care value. The dried product of female frog oviduct is used as medicine, which is called wood frog oil or husky frog oil, which is a precious animal medicinal material. The identification of this medicinal material is still mainly based on its traits, microscopic characteristics, and physical and chemical properties. However, due to the lack of obvious morphological identification characteristics and special chemical components of this medicinal material, it is still very difficult to identify Rana ovale in practice. Invention content: [0003] Aiming at the problem that it is difficult to identify the Northeast Rana and Rana egg oil in terms of traits, microscopic charac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 杨宝田
Owner SHENYANG NORMAL UNIV
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