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Method for rapidly and efficiently extracting total RNA of rice tissue

A rice, high-efficiency technology, applied in the field of total RNA extraction from rice tissue, can solve the problems of low extraction rate, long extraction time, poor integrity of total RNA, etc., and achieve the effects of high extraction efficiency and short extraction time.

Inactive Publication Date: 2010-11-03
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are many plant total RNA extraction kits on the market, but they all have certain defects, such as long extraction time, harsh extraction conditions, low extraction rate, and poor integrity of the extracted total RNA.

Method used

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  • Method for rapidly and efficiently extracting total RNA of rice tissue
  • Method for rapidly and efficiently extracting total RNA of rice tissue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Extraction of total RNA from rice root tissue:

[0022] Weigh 100 mg of the rice root tissue of Sanye Yixin, place it in a ceramic mortar (not sterilized), grind it rapidly in liquid nitrogen (within 1 min), pour it into a 1.5ml EP tube, then add 1ml of Trizol, and mix well After 15s, add 0.2ml chloroform solution and mix well for 15s. Centrifuge at 12000g for 10min, transfer the upper aqueous phase to a new 1.5ml PE tube, add 0.5ml of isopropanol, let stand at -20°C for 20min, centrifuge at 12000g for 10min, pour off the supernatant, add 1ml of 75% of Wash the precipitate twice with ethanol, take the precipitate and dry it on a clean bench, add 30 μl of 0.1% DEPC water to dissolve it.

[0023] Purity detection of total RNA in rice root tissue:

[0024] The purity of the RNA solution was detected by a Varian Cary50 UV spectrophotometer in the United States. Take 1 μl of the above-mentioned RNA solution dissolved in DEPC water, dilute it 50 times, and measure its abso...

Embodiment 2

[0032] Extraction of total RNA from rice leaf tissue:

[0033] Weigh 100 mg of the rice leaf tissue of Sanye Yixin and place it in a ceramic mortar (not sterilized), grind it rapidly in liquid nitrogen (within 1 min), pour it into a 1.5ml EP tube, then add 1ml Trizol, mix thoroughly 15s after homogenization, add 0.2ml chloroform solution and mix well for 15s. Centrifuge at 12000g for 10min, transfer the upper aqueous phase to a new 1.5ml PE tube, add 0.5ml of isopropanol, let stand at -20°C for 20min, centrifuge at 12000g for 10min, pour off the supernatant, add 1ml of 75% of Wash the precipitate twice with ethanol, take the precipitate and dry it on a clean bench, add 30 μl of 0.1% DEPC water to dissolve it.

[0034] Purity detection of total RNA in rice leaf tissue:

[0035] The purity of the RNA solution was detected by a Varian Cary50 UV spectrophotometer in the United States. Take 30 μl of the above-mentioned RNA solution dissolved in DEPC water, take 1 μl and dilute i...

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Abstract

The invention relates to a method for rapidly and efficiently extracting total RNA of rice tissue. The method is characterized by comprising the following steps of: weighing 100mg of the rice tissue, placing the rice tissue in a porcelain mortar (unsterilized), adding liquid nitrogen into the mortar and quickly grinding the rice tissue, wherein the grinding time is controlled within 1 minute; pouring the ground uniform paste into a 1.5ml EP tube, adding 1ml of Trizol, fully mixing the uniform paste and the Trizol, adding 0.2ml of solution of chloroform into the mixture and fully mixing the mixture for 15 seconds; centrifuging 12,000g of the obtained products for 10 minutes, transferring upper water phase into a new 1.5ml PE tube, adding 0.5ml of isopropyl alcohol and standing the mixture at the temperature of 20 DEG C for 20 minutes; and centrifuging 12,000g of the obtained products for 10 minutes, dumping the supernatant, adding 1ml of newly prepared 75 percent ethanol to wash the precipitate twice, drying the precipitate on a super-clean working table and adding DEPC water to dissolve the precipitate. The extraction method for the total RNA of the rice tissue has the advantages of high speed, high efficiency, completeness and the like.

Description

technical field [0001] The invention relates to a method for extracting total RNA from rice tissue in the field of biotechnology. Background technique [0002] In recent years, with the progress and development of biotechnology, gene cloning and expression research requires the extraction of high-purity RNA from plant materials. Although the extraction of RNA has become a very mature technology, it is often difficult to obtain good quality RNA in practical research. This is due to many factors (such as unreasonable preservation of materials, unclean utensils, lax operation, etc.) that cause RNA degradation. Moreover, the most suitable conditions and corresponding methods for extracting RNA from different plant materials are also different. At present, there are many plant total RNA extraction kits on the market, but all of them have certain defects, such as long extraction time, harsh extraction conditions, low extraction rate, and poor integrity of the extracted total RNA...

Claims

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Application Information

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IPC IPC(8): C12N15/10C07H21/02
Inventor 王海斌方长旬陈荣山叶陈英林志华何海斌林文雄
Owner FUJIAN AGRI & FORESTRY UNIV
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