Optimized extraction method of buckwheat RNA (ribonucleic acid)

Abstract

The invention provides an optimized extraction method of buckwheat RNA (ribonucleic acid). According to the method, cetyl trimethyl ammonium bromide is particularly adopted for optimized extraction of RNA of buckwheat roots, stems and leaves; required glassware, a gun head, an EP (epoxy resin) pipe and deionized water are subjected to high-pressure moist heat sterilization and disinfection; substances such as polysaccharides and phenols are removed by CTAB (cetyl trimethyl ammonium bromide); the phenols are removed by polyvinylpyrrolidone and beta-mercaptoethanol; an RNA precipitation effect of isopropanol is obvious; a Tris-HCl buffer solution and NaCl exerts an effect of avoiding RNA degradation. The method can extract RNA of different parts of buckwheat, such as the roots, the stems and the leaves; an extraction concentration can reach 100-500 ng / microliter; the method can be used for subsequent experiments of RT-qPCR (real-time-quantitative polymerase chain reaction), RACE (rapid-amplification of complementary deoxyribonucleic acid ends) and the like. The method is easy, simple and quick to operate and low in cost, and obtained RNA is higher in concentration and purity.

Description

technical field [0001] The invention relates to a method for extracting RNA from different parts of crop plants, in particular to an optimized extraction method for buckwheat RNA. Background technique [0002] Hexadecyltrimethylammonium Bromide (CTAB for short) is an ionic surfactant that can dissolve cell membranes and membrane proteins, depolymerize nucleoproteins, and thereby free RNA. Extraction methods can more effectively remove substances such as polysaccharides and phenols. Moreover, the extracted RNA has high concentration and purity, which can support subsequent experiments. [0003] At present, there are many kits for extracting RNA in the prior art, such as TaKaRa MiniBEST Plant RNAExtraction Kit and TaKaRa RNAiso Plus (Total RNA extraction reagent). However, the concentration and recovery of the former cannot meet the application requirements of subsequent experiments, and the cost is relatively high; the latter, although cheap, is more suitable for some herba...

AI Technical Summary

Problems solved by technology

However, the concentration and recovery of the former cannot meet the application requirements of subsequent experiments, and the cost is relatively high; the latter, although cheap, is more suitable for some herbal plants, microorganisms and animal tissue cells with less content; However, buckwheat tissue is rich in protein, phenols, and polysaccharides. At present, there is no report on a method for extracting RNA from different organs of buckwheat plants.

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