Method for rapidly extracting fungal genome DNA

Abstract

The invention discloses a method for rapidly extracting fungal genome DNA. The method comprises the following steps: adding lysate into hyphae or sporocarp which is grinded with liquid nitrogen, adding PCA, and centrifuging; adding 3M of sodium acetate into the upper-layer water phase, further adding precooled absolute ethyl alcohol, and leaving to stand for 5-10 minutes at -20 DEG C; centrifuging, abandoning the supernate, bleaching and precipitating with 75% alcohol, centrifuging, and abandoning the supernate; adding ddH2O and RNase to dissolve the components, and preserving at -20 DEG C. Compared with other methods, the method for rapidly extracting fungal genome DNA is simple to operate, high in feasibility, small in DNA extraction material, high in DNA extraction efficiency, good in integrity and stability, relatively low in impurity content, and can be directly applied to further research such as ITS sequencing. DNA of sporocarp can be also extracted, so that the hypha activation and culture time can be shortened, and an efficient and feasible molecular technical method for macro fungus molecular biological research is provided.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for rapidly extracting fungal genome DNA. technical background [0002] Macrofungi are extremely important microorganisms in the world and play important roles in people's lives. The control effect of Trichoderma on plant diseases; the metabolites of Penicillium have a good effect on the prevention and treatment of human diseases; edible fungi have important nutritional value and health care function, which can meet people's demand for healthy food, and the edible fungus industry can be transformed and utilized in large quantities Crop stalks, edible fungi are increasingly closely related to human life. For example, Poria cocos is one of the four major adjuvants of traditional Chinese medicine. Ganoderma lucidum can improve people’s immunity. Shiitake mushrooms are rich in dietary fiber and polysaccharides that can regulate human immunity. Cordyceps sinensis has...

AI Technical Summary

Problems solved by technology

The content of DNA extracted by CTAB method and SDS method is relatively high, but the purity of DNA is relatively low; the content of impurities such as phenols and sugars in DNA extracted by urea method is relatively high, which has a certain impact on subsequent research; benzyl chloride Although the extraction steps are simple and easy, the DNA quality is not stable enough. For materials with poor dispersion in the extraction buffer, the DNA extraction effect is not good; the DNA extracted by the kit method is relatively high in purity, but the DNA content is relatively high. low cost

In the extraction process of the above methods, after adding the lysate, it needs to be bathed in a water bath at 65°C for 30-60 minutes. During the water bath, it needs to be shaken every few minutes. After adding PCA, it needs to be shaken for about 10 minutes, and then let it stand. 10min, the whole process is quite time-consuming and labor-intensive

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