Method for quickly extracting ribonucleic acid (RNA) from ginseng leaf tissues

A technology of ginseng leaves and tissues, which is applied in the field of RNA extraction, can solve the problems of high drug cost, cumbersome steps, and long time consumption, and achieve the effects of protection from pollution and degradation, good extraction effect, and reduced centrifugation time

Inactive Publication Date: 2011-09-14
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The purpose of the present invention is to provide a method for rapidly extracting ginseng leaf tissue RNA in order to solve the pr

Method used

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  • Method for quickly extracting ribonucleic acid (RNA) from ginseng leaf tissues
  • Method for quickly extracting ribonucleic acid (RNA) from ginseng leaf tissues
  • Method for quickly extracting ribonucleic acid (RNA) from ginseng leaf tissues

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Experimental program
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specific Embodiment approach 1

[0019] Embodiment 1: The method for quickly extracting RNA from ginseng leaf tissue in this embodiment is carried out according to the following steps: 1. After washing and drying 200 mg of ginseng leaf tissue, grind it into powder in liquid nitrogen, and then add it to a mixture containing 750 μL of extraction buffer solution and 75 μL β-mercaptoethanol, then add 350 μL Tris to balance phenol and 350 μL chloroform, shake for 5 minutes, and then centrifuge at 13,000 r / min and 4°C for 5 minutes; 2. After centrifugation, take the supernatant and add 350 μL Tris to balance Phenol and 350 μL chloroform, shake for 5 minutes, centrifuge at 13,000 r / min, 4°C for 5 minutes, take the supernatant after centrifugation, add 750 μL of chloroform, shake for 5 minutes, and centrifuge at 13,000 r / min, 4°C for 5 minutes; 3. After centrifugation, take the supernatant, add an equal volume of isopropanol, mix well, and precipitate at -20°C for 20 minutes, then centrifuge at 13,000 r / min and 4°C fo...

specific Embodiment approach 2

[0029] Embodiment 2: This embodiment differs from Embodiment 1 in that the specification of the centrifuge tube in step 1 is 1.5ml or 2.0ml. Other steps and parameters are the same as those in Embodiment 1.

specific Embodiment approach 3

[0030] Embodiment 3: The difference between this embodiment and Embodiment 1 is that in step 3, the isopropanol should be pre-cooled before use, and the pre-cooling temperature is -20°C. Other steps and parameters are the same as those in Embodiment 1.

[0031]

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Abstract

The invention discloses a method for quickly extracting ribonucleic acid (RNA) from ginseng leaf tissues, and relates to a method for extracting the RNA. The method solves the problems that steps are complex, the consumed time is long, more medicaments are used, and the cost is high existing in the conventional method for extracting the RNA from the ginseng leaf tissues. The method comprises the following steps of: 1, grinding the ginseng leaf tissues into powder in liquid nitrogen, adding extraction buffer solution, beta-mercaptoethanol, Tris balanced phenol and chloroform, oscillating, and centrifuging; 2, taking supernatant, adding the Tris balanced phenol and the chloroform, oscillating, centrifuging, taking supernatant, adding the chloroform, oscillating, and centrifuging; 3, taking supernatant, adding isopropanol, precipitating, centrifuging, and abandoning supernatant; and 4, washing an RNA precipitate by using ethanol, and air-drying the precipitate in the air to obtain the RNA. Compared with the conventional cetyltrimethyl ammonium bromide (CTAB) method and the like, the method has the advantages that the extraction effect is higher, the time is short, and the method is easy to operate; and the extraction cost is low compared with that of a commercial kit.

Description

technical field [0001] The invention relates to a method for extracting RNA. Background technique [0002] The extraction of high-quality and complete RNA from plant tissues is the basis for transcriptome analysis, cDNA library construction, northern blot analysis, DDRT-PCR, rapid amplification of cDNA ends, RT-PCR and other molecular biology research fields. Choosing the right method for different plant materials is crucial. Because ginseng plants contain a large amount of protein, sugar, polyphenols and other metabolites of secondary metabolites, the activity of endogenous RNase is high, which seriously interferes with the extraction of RNA, resulting in the failure of the experiment. In the past, most laboratories used the traditional CTAB method and SDS method to extract plant RNA, or purchased commercial kits to extract RNA. These methods are cumbersome, time-consuming, and require many drugs and high costs. Contents of the invention [0003] The purpose of the pres...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 杨成君
Owner NORTHEAST FORESTRY UNIVERSITY
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