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Molecular biology identification method for dry sea cucumbers

A technology of molecular biology and dried sea cucumber, applied in the field of molecular biological identification of dried sea cucumber, can solve the problems of lack of scientific identification technology, inaccurate morphological identification of dried sea cucumber, etc., achieve high yield, reduce costs, and save time Effect

Inactive Publication Date: 2010-03-03
OCEAN UNIV OF CHINA
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Problems solved by technology

[0008] The purpose of the present invention is to provide a scientific, effective, simple, quick and accurate molecular biological identification method for dried sea cucumbers aimed at the inaccurate morphological identification of existing dried sea cucumbers and the general lack of scientific identification techniques

Method used

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Embodiment Construction

[0057] The present invention will be described in further detail below in conjunction with specific embodiment: the molecular biology identification method of dried sea cucumber, comprises the following steps:

[0058] (1) The high-salt method is used to extract the dried sea cucumber DNA, and the specific extraction steps are:

[0059] ① Take 100-200 mg of dried sea cucumber, soak it in Milli-Q water for 5-6 hours, and grind it into powder in liquid nitrogen;

[0060]② Transfer to a 50ml centrifuge tube, add 10ml extraction buffer, its composition is 50mM Tris, 100mM NaCl, 150mM EDTA, 1% SDS, 0.30mg / ml proteinase K, 0.1% mercaptoethanol, pH 8.0, digest at 65°C for 3h;

[0061] ③Slowly add 3ml 6mol / L NaCl, salt out to remove excess protein and polysaccharide, centrifuge at 10,000rpm at 4°C for 10 minutes, and collect the supernatant;

[0062] ④ Add phenol, chloroform, and isoamyl alcohol in an equal volume ratio of 25:24:1, centrifuge at 15,000 rpm for 10 minutes, and collect...

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Abstract

The invention relates to a molecular biology identification method for dry sea cucumbers, which comprises the following steps: extracting genome DNA of dry sea cucumbers by a high salt method; amplifying CO I genetic fragments by using a degenerate primer PCR; carrying out agarose gel electrophoresis detection on enzyme-digested products; and determining the result by the quantity and the length of electrophoretic bands. The method is used for identifying different varieties of sea cucumbers, has the characteristics of high accuracy and speed as well as convenience and is more accurate and reliable when compared with the traditional morphological identification.

Description

technical field [0001] The invention relates to an assay or inspection method comprising enzymes or microorganisms, in particular to a molecular biology identification method of dried sea cucumbers. Background technique [0002] Sea cucumbers belong to the phylum Echinodermata and the class Holothuria in taxonomy, and have great nutritional and economic value. The main species cultivated in China is Apostichopus japonicus, and dried Apostichopus japonicus is the main product purchased by consumers. With the improvement of people's living standards, the demand for sea cucumbers has increased significantly, and product identification has become a key technical problem that needs to be solved urgently. At present, my country has introduced a variety of sea cucumbers from abroad, including red sea cucumbers (Apostichopus japonicus), New Zealand sea cucumbers (Australostichopus mollis), black milk sea cucumbers (holothuria nobilis), yellow milk sea cucumbers (holothuria fuscogilv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/10
Inventor 董云伟孟宪亮纪婷婷董双林岳欣王青林田相利王芳
Owner OCEAN UNIV OF CHINA
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