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Method for simply and rapidly extracting total RNA from switchgrass tissue

A switchgrass, rapid technology, applied in the direction of recombinant DNA technology, DNA preparation, etc., can solve the problems of long time consumption, affecting the extraction efficiency of switchgrass, difficult to grind thoroughly, etc., and achieve the effect of low cost, large amount of RNA, and short extraction time

Inactive Publication Date: 2012-09-26
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Because switchgrass contains a lot of cellulose and other impurities, it is difficult to grind thoroughly during the initial grinding process of RNA extraction, and it takes too long, which affects the efficiency of switchgrass RNA extraction

Method used

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  • Method for simply and rapidly extracting total RNA from switchgrass tissue
  • Method for simply and rapidly extracting total RNA from switchgrass tissue

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A. Preparation of experimental drugs: RNA extraction solution, volume fraction is 0.1‰ DEPC water, volume fraction is 70% ethanol (prepared with 0.1‰ DEPC water).

[0026] B. Treatment of experimental supplies: Soak the utensils in DEPC water with a volume fraction of 0.1‰ overnight, then sterilize at 120°C for 80 minutes, and soak the electrophoresis tank and sampling comb in 2% hydrogen peroxide for 30 minutes.

[0027] C. Materials: Switchgrass leaves and stems

[0028] D. Extraction of total RNA:

[0029] (1) Sample grinding: Take about 0.5g of fresh switchgrass leaf or stem material in a mortar, add 3 3mm tungsten carbide grinding beads, add liquid nitrogen and grind quickly, repeat several times until it is ground into powder, and the grinding time is controlled at 1min, add the ground powder into the pre-cooled 1.5mL EP tube;

[0030] (2) Nucleic acid release: add 1mL of the prepared RNA extraction solution to the centrifuge tube in step (1), vortex fully to mi...

Embodiment 2

[0047] A. Preparation of experimental drugs: RNA extraction solution, volume fraction is 0.1% DEPC water, volume fraction is 80% ethanol (prepared with 0.1% DEPC water).

[0048] B. Treatment of experimental supplies: Soak the utensils in water with a volume fraction of 0.1% DEPC overnight, then sterilize at 120°C for 100 minutes, and soak the electrophoresis tank and sampling comb in 3% hydrogen peroxide for 50 minutes.

[0049] C. Materials: Switchgrass leaves and stems

[0050] D. Extraction of total RNA:

[0051] (1) Sample grinding: Take about 1g of fresh switchgrass leaf or stem material in a mortar, add 5 3mm tungsten carbide grinding beads, add liquid nitrogen and grind quickly, repeat several times until it is ground into powder, and the grinding time is controlled at 1min , add the ground powder into a pre-cooled 1.5mL EP tube;

[0052] (2) Nucleic acid release: add 1mL of the prepared RNA extraction solution to the centrifuge tube in step (1), vortex fully to mix ...

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Abstract

The invention provides a method for simply and rapidly extracting total RNA from switchgrass tissue, and pertains to the field of biotechnology. The concrete steps of the method comprises: a first step of preparing experimental medicines and extracting biological samples from switchgrass leaves or stems; a second step of releasing nucleic acid from the biological samples by using chemical reagents; a third step of extracting free RNA by using chloroform and precipitating the free RNA by using isopropanol to obtain the total RNA; and a fourth step of identifying the total RNA by using an ultraviolet spectrophotometer and an agarose gel electrophoresis detection method separately. The electrophoresis and data results show that the obtained RNA of plant tissues has good integrity, high purity and high yield, and can meet requirements in molecular biology research and application. Meanwhile, the method has simple experimental process, easy operation, good security, accurate results, good availability, and good repeatability.

Description

Technical field [0001] The present invention is a biotechnology field, which involves molecular biology experiments such as gene cloning and genetic function analysis of plants. It specially involves a simple and rapidly extracting total plant tissue total RNA, which is suitable for extraction of total plant tissue total RNA. Background technique [0002] With the development of modern molecular biology and its penetration to various disciplines, more and more molecular level technology is applied to plant research.Such as the use of genetically modified technology to enhance the anti -inverse of plants, improve photosynthetic efficiency, and increase biological yield.The total plant RNA extraction technology is a basic experimental technology of a plant molecular biology, and it is also a necessary prerequisite for research on plant molecular biology. It is of great significance for functional genome and genetic research.Using high-quality total RNA with high purity and good int...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 杜道林张欣黄萍薛永来杨冉田远飞
Owner JIANGSU UNIV
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