Method for extracting total RNA from root system of magnolia cylindrical

An extraction method and root technology, which are applied in the field of extraction of total RNA from the roots of Magnolia japonica, can solve problems such as inability to meet, and achieve the effects of improving extraction rate and purity, high extraction rate, and simple and easy extraction method.

Pending Publication Date: 2020-01-03
ANHUI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This method cannot meet the needs of related genetic engineering experiments of Huangshan magnolia

Method used

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  • Method for extracting total RNA from root system of magnolia cylindrical
  • Method for extracting total RNA from root system of magnolia cylindrical
  • Method for extracting total RNA from root system of magnolia cylindrical

Examples

Experimental program
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Effect test

Embodiment 1

[0072] A method for extracting total RNA from the root system of Magnolia huangshan, comprising the following steps:

[0073] 1) Dissolve 2.4g Tris in 1 / 1000 DEPC water of 100ml, add hydrochloric acid to adjust to pH=8, obtain Tris-HCl buffer solution; Dissolve 1.86gEDTA-Na 2 2H 2 O was dissolved in 100ml of DEPC water, and hydrochloric acid was added to adjust the pH to 8 to obtain EDTA-Na 2 Solution, mix the two solutions prepared above, add 4g CTAB, 4g PVP and 23.4gNaCl, mix well to obtain CTAB extract; autoclave the obtained CTAB extract at 121°C for 20min, then add CTAB Mix 2% β-mercaptoethanol and CTAB extract in the volume of the extract, use it as an extraction solvent, and place it in a water bath at 65°C to preheat; 0.5g of the fresh lateral root material of the two-year-old Huangshan Magnolia cultivated in the greenhouse Put it into a mortar pre-cooled by liquid nitrogen, and grind it fully with liquid nitrogen until any granular root system is invisible to the na...

Embodiment 2

[0084] A method for extracting total RNA from the root system of Magnolia huangshan. In the extraction process of this example, except adding LiCl solution in step 3), place it at 4° C. for 16 hours overnight; the others are the same as in Example 1, and the total RNA from the root of huangshan magnolia is finally obtained. Sample 2.

Embodiment 3

[0086] A method for extracting total RNA from the root system of Magnolia huangshan. In the extraction process of this example, step 3) added LiCl solution, and placed at 4°C for 12 hours overnight; the others were the same as in Example 1, and a total RNA sample from the root of huangshan magnolia was finally obtained. 3.

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Abstract

The invention provides a method for extracting total RNA from a root system of magnolia cylindrical. The method for extracting total RNA from the root system of the magnolia cylindrical comprises thesteps that root tissue of the magnolia cylindrical is ground into powder in liquid nitrogen, the powder is mixed with an extraction solvent, and the mixture is incubated to obtain an incubation mixedsolution; extraction treatment is performed to obtain extraction supernatant; the extraction supernatant is mixed with a lithium chloride solution, the mixture is left to stand overnight and subjectedto centrifugal treatment to obtain total RNA precipitate; the total RNA precipitate is dissolved in an SSTE buffer solution, and then the mixture is subjected to extraction treatment in a chloroform/isoamyl alcohol mixture to obtain centrifuged supernatant; the centrifuged supernatant is mixed with absolute ethanol, and the mixture is left to stand at -70 DEG C, and the mixture is subjected to centrifugal treatment to obtain precipitate; the precipitate is washed with an ethanol solution to obtain clean precipitate; and the clean precipitate is left open, DEPC-treated ultrapure water is added, and total RNA from the root system of the magnolia cylindrical is obtained.

Description

technical field [0001] The invention relates to a method for extracting RNA, in particular to a method for extracting total RNA from roots of Magnolias huangshanensis. Background technique [0002] Huangshan Magnolia (Magnolia cylindrica) is a deciduous tree of the genus Magnolia in the family Magnoliaceae. It is named for the type specimen collected from Mount Huang. Huangshan magnolia has excellent material, beautiful tree shape, and its flowers have medicinal value. It is an important garden ornamental plant and an excellent aromatic plant. It is a precious endangered species under national third-level protection. Therefore, it is of great significance to explore the germplasm resources of Magnolia magnolia at the molecular level. [0003] Huangshan Magnolia is a perennial woody plant rich in secondary metabolites such as protein, polysaccharides and polyphenols. However, polysaccharides are easy to co-precipitate with RNA, and polyphenols are easily oxidized to form qu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 杨安娜贺淑蕊孙志艳石亚冉
Owner ANHUI NORMAL UNIV
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