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Dioscorea alata lirm.sp tissue RNA extraction method

A purple-footed potato and an extraction method are applied in the field of RNA extraction, and can solve problems such as unsatisfactory effects and influence on the extraction of purple-footed potato RNA.

Inactive Publication Date: 2014-11-26
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the polyphenols and secondary metabolites in Zipai tuber are rich in polyphenols, which seriously affect the RNA extraction of Zipai tuber
At present, Trizol method, guanidine isothiocyanate one-step method (Ghawana, Paul et al. 2011; Eldh, L?tvall et al. 2012), which are widely used in China, and other reported methods for extracting total RNA from plant tissues are being applied. Satisfactory results cannot be obtained in the extraction of RNA from Zijiaban yam tissue

Method used

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  • Dioscorea alata lirm.sp tissue RNA extraction method
  • Dioscorea alata lirm.sp tissue RNA extraction method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] 1.1 Plant material

[0012] The purple-footed potato seedlings were provided by Nanchang Wancuiyuan Industrial Co., Ltd., and the roots, stems, leaves, calyx, petals, stamens and pistils of the plants were taken during flowering, and stored in liquid nitrogen.

[0013] 1.2 Main reagents and utensils

[0014] Reagents such as SDS, water-balanced phenol, imitate, isoamyl alcohol, ethanol, and DEPC used in the experiment were all purchased from Nanjing Jitian Biological Company; DNase Buffer, DNase, RNase inhibitor, etc. were all purchased from Takara Company. All configured reagents use RNase-free ddH 2 O configuration. Consumables such as pipette tips and centrifuge tubes were soaked in 0.1% DEPC water overnight and then autoclaved. The mortar and glassware were wrapped in tin foil and baked at 200 °C for more than 4 h.

[0015] 1.3 RNA extraction process

[0016] (1) Put the RNA extraction solution (2% SDS, 100mM Tris, 25mM EDTA, 2M NaCl) in a 65-degree water bath ...

Embodiment 2

[0026] Example 2 Comparison test of RNA extraction effect

[0027] 1.1 Plant material

[0028]The purple-footed potato seedlings were provided by Nanchang Wancuiyuan Industrial Co., Ltd., and the roots, stems, leaves, calyx, petals, stamens and pistils of the plants were taken during flowering, and stored in liquid nitrogen.

[0029] 1.2 Main reagents and utensils

[0030] Reagents such as SDS, water-balanced phenol, imitate, isoamyl alcohol, ethanol, and DEPC used in the experiment were all purchased from Nanjing Jitian Biological Company; DNase Buffer, DNase, RNase inhibitor, etc. were all purchased from Takara Company. All configured reagents use RNase-free ddH 2 O configuration. Consumables such as pipette tips and centrifuge tubes were soaked in 0.1% DEPC water overnight and then autoclaved. The mortar and glassware were wrapped in tin foil and baked at 200 °C for more than 4 h.

[0031] 1.3 RNA extraction process:

[0032] In order to illustrate the advanced nature...

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PUM

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Abstract

The invention belongs to the field of biotechnology and discloses a dioscorea alata lirm.sp tissue RNA extraction method. Based on a SDS method, in material crushing, water-insoluble polyvinylpyrrolidone (PVPP) is used for preventing phenol substance oxidation, a high-salt solution is used for removing polysaccharide pollution and 10% of KAC (having pH value of 5.2 and a concentration of 3M) and 250% of ethanol are used for RNA deposition. The RNA extracted by the dioscorea alata lirm.sp tissue RNA extraction method has good purity and integrity, is not polluted by impurities such as DNA, proteins and polysaccharides and is convenient for downstream molecular biology experiments.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to an RNA extraction method. Background technique [0002] Purple foot plate yam is a perennial twining vine of the family Dioscoreaceae (Dioscoreaceae). It is named for its foot-shaped potato and purple flesh. Purple foot potato is rich in protein, amino acid, vitamin, polysaccharide, trace elements and dietary fiber. It is a health food integrating food and medicine. The development of starch sugar technology has a huge market in China, and it can be transformed into industrial raw materials such as alcohol, vinegar, and syrup through modern bioengineering technology [2] , and Zijiao yam has gradually emerged due to its high yield (up to several tons per mu) and high starch content, and is expected to become a "super crop" that combines food, medicine and energy. [0003] With the recognition of its importance, the research on purple footed potato has also been widely carried out. Amo...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 罗火林杨柏云罗丽萍熊冬金黄学勇
Owner NANCHANG UNIV
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