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Method for extracting total RNA from sandalwood wood tissue

A wood and tissue technology, applied in the field of molecular biology, can solve problems such as difficulty in extracting total RNA from sandalwood wood tissue, and achieve the effects of strong promotion and application value, strong practicability, and good integrity.

Inactive Publication Date: 2016-05-25
SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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  • Abstract
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Problems solved by technology

[0004] The purpose of the present invention is to provide a kind of high-quality method for extracting the total RNA of sandalwood tissue for the deficiency that the total RNA of sandalwood tissue is difficult to extract in the prior art

Method used

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  • Method for extracting total RNA from sandalwood wood tissue

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Embodiment 1

[0020] The following plastic centrifuge tubes and pipette tips used in the method of extracting total RNA from sandalwood wood tissue were soaked in chloroform for 30 minutes, then sterilized under high pressure at 121°C for 30 minutes, and dried for later use; mortar and metal spoons were used with tin foil Wrap in paper and bake in an oven at 180°C for 3-5 hours. DEPC water is deionized double-distilled water with a final concentration of 0.1% DEPC treated and sterilized by high temperature and high pressure.

[0021] Guanidine isothiocyanate extract, its formula is: 4mol / L guanidine isothiocyanate, 2mmol / LTris-HCl (pH8.0), 1% β-mercaptoethanol (v / v), solvent is water, and its preparation The method is to mix the above-mentioned ingredients evenly, and then sterilize it for later use.

[0022] SDS buffer solution, its formula is: 0.2%SDS (w / w), 100mmol / LNaCl, 50mmol / LTris-HCl (pH=8.0), 10mmol / LEDTA (pH=8.0), solvent is water, and its preparation method is to After the abov...

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Abstract

The invention discloses a method for extracting total RNA from sandalwood wood tissue. The method comprises the following steps: grinding taken sandalwood wood tissue into powder, adding the powder to a guanidinium isothiocyanate extract solution, and centrifuging after an ice bath; taking supernatant, adding an SDS buffer solution for mixing evenly, adding water-saturated phenol / chloroform / isoamyl alcohol for mixing evenly, and centrifuging the mixture; taking the supernatant, adding chloroform for mixing evenly and centrifuging the mixture; taking the supernatant, adding absolute ethyl alcohol and sodium acetate solution sediment RNA, and centrifuging the mixture for removing the supernatant; and taking the sediment and washing the sediment with ethanol to obtain the total RNA. The ratio of OD260 to OD280 of the extracted total RNA of the sandalwood wood tissue is 1.8 to 2.0; the ratio of OD260 to OD230 is 2.0 to 2.3; the concentration of the RNA water solution is 350.0-500.0ng / mu L; electrophoresis detection shows that the RNA strip is clear and good in integrity; and the brightness at 28S is 1.9-2.1 times that at 18S. The total RNA of the sandalwood wood tissue extracted by the method provides a good foundation for related researches of sandalwood molecular biology, and the method has relatively high popularization and application values.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular to a method for extracting total RNA from sandalwood wood tissue. Background technique [0002] Sandalwood (Santalum album L.) is a semi-parasitic evergreen tree of the genus Santalum of the family Santalaceae. It is a valuable and rare plant, mainly distributed in India, Indonesia, Australia and some islands in the Pacific Ocean. Humans have used it for thousands of years, and it is especially famous because its wood can be used for carving and refining sandalwood oil, known as "green gold". There is no native distribution of sandalwood in China, and its finished or semi-finished products have been imported from abroad in the past. In 1962, the South China Botanical Garden of the Chinese Academy of Sciences introduced sandalwood seeds from Indonesia to the mainland for the first time and succeeded in breeding. Later, it also successfully introduced the high-quality provenance of "...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 张新华牛美云程庆伟马国华
Owner SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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