Method for extracting wheat total RNA

A wheat and centrifuge tube technology, applied in the field of plant biology, can solve problems such as unsatisfactory extraction results, and achieve the effects of reducing pollution levels, shortening extraction time, and avoiding interference.

Inactive Publication Date: 2020-02-07
漯河市农业科学院
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Problems solved by technology

[0008] The purpose of the present invention is to provide a method for extracting wheat total RNA, which solves the problem that there are ma

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  • Method for extracting wheat total RNA

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[0034] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work shall fall within the protection scope of the present invention.

[0035] See figure 1 , A method for extracting total RNA from wheat, including the following steps:

[0036] Step 1: Take fresh or preserved wheat tissue in a refrigerator at -70°C, add liquid nitrogen in a mortar and grind it with a pestle to powder; add liquid nitrogen during the grinding process to keep the powder in a frozen state; obtain wheat powder;

[0037] Step 2: Move the wheat powder to a 1.5ml centrifuge tube A, add 1ml of ice-co...

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Abstract

The invention discloses a method for extracting wheat total RNA. The method comprises the following steps of step 1, taking fresh wheat tissue or wheat tissue which is preserved in a refrigerator of minus 70 DEG C, and adding liquid nitrogen in a grinding process to enable powder to maintain a frozen state so as to obtain wheat powder; step 2, transferring the wheat powder into a 1-5ml centrifugaltube A; putting the centrifugal tube A on ice to obtain wheat sample mixed liquor A; step 3, adding a chloroform solution of which the volume is the same as that of the wheat sample mixed liquor A tothe wheat sample mixed liquor A obtained in the step 2, and during adding, performing uniform mixing for 2-3 times so as to obtain wheat sample mixed liquor B; step 5, adding isopropyl alcohol of which the volume is 0.25 time of that of a centrifugal tube B and RNA lysate of which the volume is 0.25 time of that of the centrifugal tube B to the centrifugal tube B, adding clear liquor obtained inthe step 4 into the centrifugal tube B until the centrifugal tube B is full, performing sufficient and uniform mixing to obtain a mixture, and enabling the mixture to be kept on the ice for 30min; andstep 7, adding an appropriate amount of water treated with 0.1% DEPC, performing dissolving, and collecting solute namely the wheat tissue RNA. According to the method disclosed by the invention, theusage of sodium acetate and acid phenol is removed, and experiment results indicate that the yield and the quality of the RNA cannot be reduced.

Description

technical field [0001] The invention relates to the field of plant biotechnology, in particular to a method for extracting wheat total RNA. Background technique [0002] RNA with good integrity and high purity is the premise and basis for molecular biology research such as wheat RT-PCR, Northern blot analysis, and cDNA library construction. Wheat tissue cells are rich in secondary substances such as polysaccharides, lipids, polyphenols, and pigments, which are difficult to remove during RNA extraction. The presence of polysaccharides and lipids will not only reduce the solubility of RNA, but also inhibit the activity of many tool enzymes; secondary substances such as polyphenols and pigments are easily oxidized during the extraction process and cause RNA to brown, affecting the use of RNA for further processing. Therefore, whether these interfering substances can be removed as much as possible during the extraction process is the key to obtaining high-quality wheat total RN...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 曹燕燕李雷雷王君张璐郭春强葛昌斌黄杰乔冀良张振永齐双
Owner 漯河市农业科学院
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