Extraction method of santalum album high-content essential oil tissue total RNA

A high-content, tissue technology, applied in the field of molecular biology, can solve the problems of difficult RNA extraction, low yield, low quality, etc., to achieve the effect of clear RNA bands, simplified operation steps, and improved yield.

Active Publication Date: 2020-05-19
SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Based on the wood tissue of sandalwood with high content of essential oil, polysaccharide, polyphenol and other secondary metabolites, the present invention develops

Method used

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  • Extraction method of santalum album high-content essential oil tissue total RNA
  • Extraction method of santalum album high-content essential oil tissue total RNA
  • Extraction method of santalum album high-content essential oil tissue total RNA

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Embodiment 1

[0026] The following plastic centrifuge tubes and pipette tips used in the method of extracting total RNA from the heartwood tissue of sandalwood stems were soaked in DEPC water for 30 minutes, then sterilized under high pressure at 121°C for 30 minutes, and dried for later use; mortar and metal spoon Wrap it in tin foil and bake in an oven at 180°C for 4-6 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.

[0027] Tris-HCl extract, its formula is: 100mM Tris-HCl, 10mM EDTA, 50mM lithium dodecyl sulfate (LSD), 200mM lithium chloride (LiCl), 1% (g / ml) Tergitol Nonidet P-40 ( NP-40)], 10mM dithiothreitol (DTT), 3% (g / ml) polyvinylpolypyrrolidone (PVPP), pH 8.0, all reagents are configured with a certain concentration of mother liquor with DEPC water, and sterilized for subsequent use; Mix evenly in a certain proportion.

[0028] The formula of TE buffer solution is: 10mM T...

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Abstract

The invention discloses a method for extracting santalum album high-content essential oil tissue total RNA. The method for extracting the santalum album high-content essential oil tissue total RNA comprises the following steps: grinding santalum album wood tissue fluid in liquid nitrogen so as to obtain powder; adding the powder into Tris-HCl extraction liquid, carrying out freezing treatment at -80 DEG C, and carrying out centrifuging so as to obtain liquid supernatant; adding sodium acetate and isopropanol into the liquid supernatant so as to have RNA precipitated, carrying out centrifuging,and dissolving the precipitates by using a TE buffer solution; adding cetyltrimethylammonium bromide into the dissolved precipitates, carrying out uniform mixing, carrying out extraction by using chloroform / isoamyl alcohol, and centrifuging the extract so as to obtain liquid supernatant; and then, adding lithium chloride into the liquid supernatant so as to carry out RNA precipitation overnight, centrifuging the precipitated RNA, and washing the centrifuged RNA by using ethanol so as to obtain the total RNA. Being tested by using Nanodrop, the santalum album wood tissue total RNA extractedby the method disclosed by the invention has an OD<260> / OD<280> ratio of 2.0 or above, an OD<260> / OD<230> ratio of 2.3 or above, an RNA concentration in aqueous solution of 800.0 ng/[mu]L or more,and RIN higher than 7.5-1; and 1.0% denaturing agarose gel electrophoresis shows that the RNA strip is clear and good in integrity, so that the santalum album wood tissue total RNA has the advantagesof being high in quality and high in concentration.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a method for extracting total RNA from tissues with high content of sandalwood essential oil. Background technique [0002] Sandalwood (Santalum album L.) is a semi-parasitic evergreen tree of the genus Santalum in 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 ...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 张新华张婷陈晓宏郑枫曾宋君马国华
Owner SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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