Method for extracting total ribonucleic acid (RNA) from bracts and inflorescences of Anthurium andraeanum

An extraction method, the technology of Anthurium, applied in the field of plant total RNA extraction, can solve the problems of total RNA extraction and different applicability, and achieve the effects of low cost, easy preparation, and harm reduction

Inactive Publication Date: 2012-07-18
FLOWERS & PLANTS RES DEV CENT ZHEJIANG ACAD OF AGRI SCI
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Problems solved by technology

[0003] The ornamental organ of anthurium is its bracts, not flowers, but conventional hybrid breeding must study its floral organs, so the total RNA extraction of anthurium bracts and inflorescences is very necessary in the process of molecular biology breeding, but currently foreign The research on the molecular biology of anthurium has

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  • Method for extracting total ribonucleic acid (RNA) from bracts and inflorescences of Anthurium andraeanum
  • Method for extracting total ribonucleic acid (RNA) from bracts and inflorescences of Anthurium andraeanum
  • Method for extracting total ribonucleic acid (RNA) from bracts and inflorescences of Anthurium andraeanum

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Embodiment 1

[0023] 1. Preparation of reagents

[0024] 1) Preparation of initial RNA extraction solution: Weigh 2g CTAB, 2g PVP, 0.73g EDTA, 11.68g sodium chloride in a clean 250ml Erlenmeyer flask, add water to 100ml, then add 0.1% DEPC ( 0.1ml), shake to mix the solution, and let it stand at room temperature for 1~2d in an airtight state, then 1kg / cm 2 Steam sterilize at 120°C for 20 minutes, add 1.21g Tris under aseptic conditions after cooling, and adjust the pH value to 8.0 with 100% concentrated hydrochloric acid in mass concentration, seal it for later use, add β-mercaptoethanol (2ml ).

[0025] 2) Preparation of 8M LiCl solution: Dissolve 24.16g lithium chloride in 50ml distilled water and add 50μl DEPC at the same time, mix well and let it stand at room temperature for 1-2 days, then 1kg / cm 2 Steam sterilize at 120°C for 20 minutes, and cool for later use.

[0026] 3) Preparation of 0.5mol / L EDTA (pH8.0): 18.6g EDTANa 2 ·H 2 O was dissolved in distilled water, and the volume...

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Abstract

The invention discloses a method for extracting total ribonucleic acid (RNA) from bracts and inflorescences of Anthurium andraeanum. The method comprises the following steps of: grinding young and tender bracts and inflorescences of the Anthurium andraeanum by using liquid nitrogen, standing an RNA extracting solution in a water bath at the temperature of 63 to 67 DEG C for 5 to 8min, adding chloroform A in an amount which is one fifth of the volume of the RNA extracting solution, violently shaking for 15 to 20s again, standing for 5 to 8min at room temperature, centrifuging to obtain a supernatant, treating the supernatant by using chloroform and a 8M LiCl solution, and centrifuging to obtain the total RNA of the bracts and the inflorescences of the Anthurium andraeanum. By the method, the using amount of toxic and harmful medicines is furthest reduced, and the hazard of RNA extraction on human bodies is reduced; high-quality RNA which is adequate and pure and is not subjected to deoxyribonucleic acid (DNA) contamination can be obtained; and the method is easy to operate, and a high-quality product is obtained in a short time.

Description

(1) Technical field [0001] The invention relates to a method for extracting plant total RNA, in particular to a method for extracting total RNA from anthurium bracts and inflorescences. (2) Background technology [0002] Anthurium andraeanum (Anthurium andraeanum) is a world-renowned potted and cut flower with a huge market share and a wide range of planting areas in my country. With the expansion of applications, the breeding of anthurium varieties is in the ascendant, but due to the limitation of resources, it is becoming more and more difficult to breed new varieties, so breeding experts all over the world are pinning the cultivation of new varieties on biotechnology. Biotechnology breeding based on the development and growth of plant molecular biology must go deep into the nucleic acid level. During this period, obtaining reliable and rich nucleic acid information is a prerequisite for good breeding work, and RNA extraction is almost an essential part. [0003] The orna...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 马广莹朱开元邹清成刘慧春周江华
Owner FLOWERS & PLANTS RES DEV CENT ZHEJIANG ACAD OF AGRI SCI
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