Method for extracting total RNA (Ribonucleic Acid) from woody mangrove plants and special extracting solution thereof

A technology of mangrove plants and plants, applied in the direction of DNA preparation, recombinant DNA technology, etc., can solve problems such as improper handling, RNA yield impact, and biological experiments, and achieve the effect of preventing oxidation

Inactive Publication Date: 2011-08-03
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, improper handling of polysaccharides will lead to two consequences in the extraction of RNA, which will affect further biological experiments.
First, a large amount of RNA is also removed while remo

Method used

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  • Method for extracting total RNA (Ribonucleic Acid) from woody mangrove plants and special extracting solution thereof
  • Method for extracting total RNA (Ribonucleic Acid) from woody mangrove plants and special extracting solution thereof
  • Method for extracting total RNA (Ribonucleic Acid) from woody mangrove plants and special extracting solution thereof

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Embodiment 1

[0032] Example 1. Using the method of the present invention to extract plant total RNA

[0033] 1. Using the method of the present invention to extract RNA

[0034] Extraction buffer: composed of Tris-HCl, EDTA, NaCl, SDS, DTT, β-mercaptoethanol and water; the final concentration of Tris-HCl is 100mmol·L -1 , The final concentration of EDTA is 500mmol·L -1 , The final concentration of NaCl is 500mmol·L -1 , The final concentration of SDS is 2% (mass percentage), and the final concentration of DTT is 5mmol·L -1 The final concentration of β-mercaptoethanol is 1.5% (volume percentage); the pH is 8.5. Take 0.5ml extraction buffer and put it in a 1.5ml centrifuge tube to preheat at 65°C.

[0035] The RNAs of Kadelia candel, Rhizophora stylosa and Lumnitzera racemosa were extracted separately. The methods are as follows:

[0036] 1. Place 0.2g of young leaves in liquid nitrogen, add PVP (molecular weight 40,000) (the amount of PVP added is 1-2% of the leaf mass), grind to powder, quickly tr...

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Abstract

The invention discloses a method for extracting total RNA (Ribonucleic Acid) from woody mangrove plants and special extracting solution thereof. The extracting solution provided in the invention comprises Tris-HCl, EDTA (Ethylene Diamine Tetraacetic Acid), NaCl, SDS (Sodium Dodecyl Sulfonate), DTT (Dithiothreitol), beta-mercaptoethanol and water, wherein the final concentration of the Tris-HCl is 75-125 mmol/L, the final concentration of the EDTA is 400-600 mmol/L, the final concentration of the NaCl is 400-600 mmol/L, the final concentration of the SDS is 1%-3% (in weight percent), the final concentration of the DTT is 3-8 mmol/L, and the final concentration of the beta-mercaptoethanol is 1%-3% (in weight percent); and the pH value is 8-9. The method provided in the invention is used for extracting the total RNA from the woody mangrove plants by using the extracting solution. The RNA extracted by the method can be directly used for synthesizing cDNA (complementary Desoxyribonucleic Acid) and carrying out molecular biology operations, such as library establishment, molecular clone and the like.

Description

Technical field [0001] The invention relates to a method for extracting total RNA of woody mangrove plants and a special extracting solution thereof. Background technique [0002] Mangrove (Mangrove) is a woody plant community on the tropical coast. It is a tropical and southern subtropical coastal plant community. It is named because it is mainly composed of plants of the mangrove family. Mangroves are characteristic plants of wetlands. There are 61 species worldwide: Kadelia candel (Kadelia candel) and Rhizophora stylosa (Rhizophora stylosa) of the mangrove family, and Lumnitzera racemosa (Lumnitzera racemosa) of the Junzi family. [0003] It is a precious species resource with the unique characteristics of the ecosystem on the edge of the sea and land. With the rapid development of industry and agriculture in modern river basins, the rapid growth of population and economy in coastal cities, and human disturbance, environmental pollution in the estuary bay area has become increa...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 何振艳麻密陈宏林
Owner INST OF BOTANY CHINESE ACAD OF SCI
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