Method for extracting wheat total RNA

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...

AI Technical Summary

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 many methods for extracting wheat total RNA at present, but the extraction effect of these methods is not very satisfactory in actual operation.

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