Method for extracting total RNA of microalgae cells

Abstract

The invention discloses a method for extracting the total RNA of microalgae cells. The method has the advantages that microalgae is placed in a plant RNA extraction auxiliary fluid containing silica sand to be grinded, thereby achieving that grinding and polysaccharide removal treatment of the microalgae is rapidly completed through one step, greatly reducing the demands for the microalgae thallusamount, greatly simplifying the experimental operation, reducing the experimental time consumption. According to the method, breaking of the microalgae cell wall is combined with the removal of the microalgae so that the cell wall breaking and polysaccharide removal are completed within 5 min; through the innovation and improvement of the whole extraction method, the extraction time of the microalgae RNA is shortened to less than 30 min; on this basis, it is achieved that all the operations of the extraction of the microalgae RNA can be carried out in a general laboratory environment, and a special extraction environment of RNA enzyme removal is not needed; all experimental supplies do not need DEPC water for RNA enzyme removal treatment; the DEPC water is not harmful to the health of researchers while the early-stage preparation work is reduced, and the safety of experimenters is protected.

Description

technical field [0001] The invention belongs to the field of RNA extraction, and in particular relates to a method for extracting total RNA from microalgae cells. Background technique [0002] Microalgae is a general term for a class of tiny organisms that contain chlorophyll and can perform photosynthetic autotrophic growth. Because its cells are rich in nutrients such as algal polysaccharides, proteins, and fatty acids, and it has the characteristics of rapid growth through photosynthesis, it has attracted widespread attention and has broad application prospects in food, health care, and medical fields. Due to its high photosynthetic efficiency, short growth cycle, high oil content, and the physical and chemical properties of microalgae biodiesel similar to traditional diesel, microalgae has become the third generation of biodiesel, and is the most promising biodiesel for industrialization. . [0003] Although microalgae has high value and good application prospects, the...

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Problems solved by technology

[0004] 1. The sample volume required for microalgae RNA extraction is large: the first step of traditional microalgae RNA extraction is usually liquid nitrogen grinding, so the large sample volume makes it difficult to dynamically monitor the level of microalgae RNA

[0005] 2. Microalgal polysaccharides seriously affect RNA extraction: algal cells are rich in algal polysaccharides, and polysaccharides will precipitate together with RNA after the microalgae is broken, which seriously reduces the quality of RNA extraction, and the content of polysaccharides in the later stage of microalgae culture increase, which increases the difficulty of RNA extraction

[0006] 3. Hazards of DEPC water: Generally, when extracting RNA, all experimental supplies and laboratory facilities must be treated with DEPC water to remove RNase, and DEPC water is carcinogenic, which seriously affects the personal safety of researchers.

[0007] 4. It takes a long time to extract microalgae RNA: due to the influence of factors such as the removal of RNase from all experimental supplies and workbenches for extracting microalgae RNA, the liquid nitrogen grinding of microalgae, and the time-consuming basis of Trizol extraction of RNA, the general The RNA extraction method from microalgae takes a long time, which will affect the yield and integrity of RNA on the one hand, and on the other hand, it is not conducive to the smooth development of the overall experiment.

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