Method for extracting total RNA from leechee

Abstract

The invention discloses a method for extracting total RNA from leechee, which comprises: grinding a leechee leaf sample treated by liquid nitrogen, and extracting with acetone under vacuum; adding RNA extraction solution into a plant material, which is subjected to vacuum extraction, for extraction, centrifuging, precipitating supernate with LiCl, centrifuging and collecting precipitate; washing and precipitating by using LiCl solution, centrifuging and discarding the supernate; and precipitating, drying, dissolving in diethyl pyrocarbonate (DEPC) sterilizing solution, precipitating and storing at 80 DEG C below zero for later use. Compared with the conventional extraction method, the method is simple in operation, low in extraction cost and high in repeatability and adaptability, can acquire high-quality and high-concentration RNA, and lays a foundation for further molecular biological research. Meanwhile, the method can also be used for extracting total RNA from the plant tissues of other species rich in polysaccharide and polyphenol substances.

Description

technical field [0001] The invention relates to molecular biology experiments such as gene cloning and gene function analysis of litchi, in particular to a method for extracting total RNA from litchi, which is suitable for extracting total RNA from plant tissues rich in polysaccharides and phenolic compounds such as litchi. Background technique [0002] Litchi is a well-known characteristic fruit tree in tropical and subtropical regions of my country, and its research has attracted widespread attention. As far as litchi genetics research is concerned, the traditional research is mainly carried out from the aspects of morphology and plant physiology, and has gradually turned to the use of molecular biology methods to carry out molecular research on it. [0003] Plant total RNA extraction technology is a basic experimental technique of plant molecular biology and a necessary prerequisite for plant molecular biology research. The isolated high-quality total RNA with high purit...

AI Technical Summary

Problems solved by technology

Commonly used methods for extracting total RNA from plant tissues include: guanidine method, phenol method, Tris-boric acid method, CTAB method, TRIZOL kit method, etc. These methods have been widely used in the extraction of total RNA from many plant tissues, but some plant tissues The inclusions in RNA are extremely complex, especially when they are rich in polysaccharides, polyphenols, tannins, and some unidentified secondary metabolites, which are not conducive to the isolation and purification of RNA, so whether they can be effectively removed or inhibited The interference of metabolites such as polysaccharides and phenols is the key to the success of extracting high-quality plant RNA

[0004] In intact cells, polyphenolic compounds, polysaccharides, proteins, anthocyanins, etc., and some unknown secondary metabolites are spatially separated from nucleic acids, but when tissues are ground and cells are broken, these substances will be separated from nucleic acids. RNA interaction: polysaccharides form insoluble jelly and co-precipitate with RNA; phenolic compounds are easily oxidized to brown substances, and then irreversibly combine with RNA, resulting in loss of RNA activity and RNA loss when extracted with chloroform Lost, or form insoluble complexes

Images