Rapid extraction method of Fagopyrum tataricum grain RNA

Abstract

The invention provides a rapid extraction method of Fagopyrum tataricum grain RNA. The method comprises the following steps: (1) disinfection treatment is carried out; (2) pyrolysis with CTAB extractis carried out; wherein a formula of the CTAB extract is as follows: 2% CTAB (W / V), 60mM of Tris-HCl, 25mM of EDTA, 15% absolute ethyl alcohol, pH value is 8.0; (3) 12000rpm centrifugation is carriedout at a normal temperature for 5-10 minutes; (4) chloroform-isoamyl alcohol mixed liquor is added into a supernatant, after well mixing, potassium acetate whose pH value is 4.6 is added, 12000rpm centrifugation is carried out at a normal temperature for 5-10 minutes, and the supernatant is taken; (5) a LiCl solution is added into the supernatant, standing and deposition is carried out for 1-2 hours at -20 DEG C, precipitate is taken and 12000rpm centrifugation is carried out at 4 DEG C for 10 minutes, precipitate at the tube bottom is rinsed with ethanol, redistilled water is added for dissolving, and electrophoresis detection is carried out. The method is carried out in order to obtain RNA of high purity, and the method has the advantages of less RNA losses, high extraction concentration, and fast extraction speed; after electrophoresis detection, bands are distinct, RNA has good integrity, OD260 / OD280 ratio is 1.8-2.0, and the method can be used for analysis of gene expression or molecular clone and other molecular biology researches.

Description

technical field [0001] The invention belongs to the technical field of biological genes, and in particular relates to a method for quickly extracting tartary buckwheat grain RNA. Background technique [0002] Tartary buckwheat, also known as tartary buckwheat, belongs to the genus Buckwheat of Polygonaceae. It is a kind of medicinal and edible crop. The active ingredients of tartary buckwheat are rich in flavonoids, phenolic compounds, inositol, weakly alkaline starch and active protein. , is a food that can be eaten as a meal, and its nutrient content is rich. It is rich in selenium itself, which can naturally supplement selenium to the human body. It has excellent nutritional and health care value and extraordinary therapeutic effect. The planting area and output of tartary buckwheat in my country rank first in the world, and it mainly grows in the southwest mountainous area of ​​China. Tartary buckwheat tea in Sichuan is also widely spread. [0003] The extraction of RNA...

AI Technical Summary

Problems solved by technology

However, since plant cells have a hard cell wall, and plant cells contain polyphenols, polysaccharides, terpenoids, or other unidentified secondary metabolites, these substances and nucleic acids are separated from each other in intact cells, but when the tissue is Grinding, after the cells are broken, these substances will interact with the RNA, so it is extremely difficult to isolate high-quality RNA from plant tissues

[0004] Chinese patent document CN 106967711 A discloses an optimized extraction method of buckwheat RNA, which uses CTAB extraction buffer solution to extract RNA from different parts of buckwheat roots, stems and leaves, which can remove polysaccharides and phenols and other substances, and obtain a higher concentration However, this method uses a high-salt buffer system, which dissolves the DNA well and also causes some RNA loss, and it uses the traditional chloroform-isoamyl alcohol extraction step, which requires multiple steps Repeated extraction can obtain high-purity RNA, which is not only time-consuming and labor-intensive, but also easily causes RNA loss, and the extraction efficiency is low, and after repeated centrifugation at 4°C, the lysate tends to become viscous and difficult to absorb. High pressure moist heat sterilization takes a long time

In addition, because the same method has different extraction effects in different plant tissues; even the RNA extraction methods of different tissues of the same plant will be very different, so the method of CN 106967711A is not suitable for the extraction of tartary buckwheat grains , the extraction effect is poor

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