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Method for quickly extracting low molecular weight RNA of plant

A low molecular weight, extraction method technology, applied in sugar derivatives, organic chemistry, etc., can solve problems such as degradation, and achieve the effect of good integrity, high purity, simple and fast experimental process

Inactive Publication Date: 2010-06-16
RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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Problems solved by technology

The conventional method is to extract relatively pure molecular weight RNA through fractionation and several precipitations, and there is a risk of degradation

Method used

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Examples

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Embodiment Construction

[0020] Below in conjunction with embodiment the present invention is further described.

[0021] A rapid extraction method for plant low molecular weight RNA, the extraction buffer ratio is: 2% PVP, 2% CTAB, 0.1M to 0.6M sodium acetate, 25mM EDTA, 2% β-ME, the pH value range of the extraction buffer It is 4.0-8.0.

[0022] The plant tissues used in this example were fresh, young papaya, Arabidopsis, cassava and rubber tree leaves.

[0023] The extraction method of low-molecular-weight RNA in plants is realized according to the following steps:

[0024] ①Preheat the extraction buffer at 65°C for 30 minutes;

[0025] ②Take fresh plant tissue samples and quickly grind them into powder in liquid nitrogen;

[0026] ③According to the ratio of 0.1g fresh sample per 1mL of extraction buffer, quickly transfer the ground sample powder to the extraction buffer, and place it in a water bath at 65°C for 20 minutes, during which it is shaken continuously;

[0027] ④ After cooling on ice...

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Abstract

The invention discloses a method for quickly extracting low molecular weight RNA of plant. After cells are disrupted, genomic DNA and high molecular weight RNA form a DNA-protein complex and a RNA-protein complex with protein respectively; and the DNA-protein complex and the RNA-protein complex are insoluble in acid and low-salt environment, and can be removed and extracted through acid and phenol. The low molecular weight RNA stays in supernatant solution, and can be recovered through the precipitation of ethanol or isopropanol. The method is quick and efficient, and can recover the low molecular weight RNA only by one-step precipitation after phenol extraction. The low molecular weight RNA extracted by the method has high purity, and can be used for the cloning of the low molecular weight RNA and the detection of Northern blot hybridization. The method has simple and quick experimental process, and is suitable for the extraction of the low molecular weight RNA of various plant tissues.

Description

technical field [0001] The invention relates to a method for extracting low-molecular-weight RNA from plants, in particular to a rapid extraction method for low-molecular-weight RNA, which is suitable for the extraction of low-molecular-weight RNA from most plant tissues. Background technique [0002] At present, low molecular weight RNA (low molecular weight RNA, LMW RNA) refers to a class of RNA molecules with a small molecular weight and a small number of bases in cells, mainly including small interfering RNAs with a length of 20-24 nucleotides ( siRNA), microRNA (miRNA), transfer RNA (tRNA) and 5S RNA, etc. These RNAs play a very important role in the process of cell activity, especially the regulation of gene expression by siRNA and miRNA is a current research hotspot. [0003] The extraction of low molecular weight RNA is a basic work of his research. Conventional low-molecular-weight RNA extraction methods are generally divided into two steps: first isolate total RN...

Claims

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Application Information

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IPC IPC(8): C07H21/02
Inventor 程汉安泽伟黄华孙高静
Owner RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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