Method for efficient extraction of total RNA in rice planthopper
A high-efficiency technology for rice planthoppers, applied in the field of agricultural biology, can solve the problems of ineffective and efficient extraction, and achieve the effects of significant practical value, good integrity, and simple and convenient methods
Inactive Publication Date: 2015-11-25
ZHENJIANG AGRI SCI INST JIANGSU HILLY AREAS
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Problems solved by technology
[0006] Purpose of the invention: In order to overcome the problem that the total RNA in the wings of rice planthoppers cannot be effectively and efficiently extracted in the prior art, the present invention proposes a method that can obtain high-quality RNA without being degraded to the minimum by exogenous or endogenous RNases. The method for efficiently extracting total RNA in rice planthopper wings that can maintain the purity of the extracted total RNA to the maximum extent to facilitate the use of subsequent experiments
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[0030] The present invention will be further described in detail below in conjunction with the embodiments:
[0031] 1) Test steps:
[0032] The brown planthopper was selected as the test object.
[0033] a) Sterilization of experimental equipment and consumables and RNase inactivation treatment: Use 0.1% DEPC aqueous solution to soak the experimental equipment and pipette tips for more than 24 hours. Some consumables are sterilized by high temperature, high pressure, high pressure and humid heat in a sterilizer, and glass rods and quartz sand are used for drying. Dry heat sterilization at 200°C for more than 2 hours; wipe clean the work surface with RNase scavenger; soak the dissecting needle in absolute ethanol and burn and inactivate it with an alcohol lamp;
[0034] b) Separation of brown planthopper wings: Use a straw to suck a certain number of brown planthoppers into a centrifuge tube and quickly place them in a liquid nitrogen tank for tens of seconds to make them die quickly....
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The invention discloses a method for efficient extraction of total RNA in rice planthopper. Compared with conventional means of extracting RNA by virtue of Trizol, the method has the following advantages: the method is simple and convenient, and is capable of effectively extracting total RNA in planthopper with the addition of freeze-storage and quartz sand aid-grinding in steps; the extracted total RNA in the planthopper is not degraded to the minimum extent through endogenous or exogenous RNA enzyme, and the purity of the extracted total RNA is kept to the greatest extent, so as to facilitate the utilization of subsequent tests; the obtained planthopper total RNA is high in purity and good in completeness, and the RNA, with the concentration, can meet the demands of various experiments; and the total RNA is applicable to operation application of molecular biology in reverse transcription cDNA, PCR, qPCR and like downstream operations, and the total RNA is significant in practical value.
Description
Technical field [0001] The invention relates to the field of agricultural biotechnology, in particular to a method for efficiently extracting total RNA from the wings of rice planthoppers. Background technique [0002] Nilaparvatalugens, Laodelphaxstriatellus and Sogatellafurcifera belong to the order Hemiptera and Delphacidae, and are currently important pests on rice in Asia. It not only causes adults and nymphs to swarm and suck the phloem sieve juice of rice, which affects the growth and development of rice, but it is also a vector insect for the transmission of a variety of virus diseases. The incidence of black-streaked dwarf in southern rice is related to the migration of venomous white-backed planthoppers. Adult rice planthoppers can be divided into long-winged and short-winged types. The wing differentiation of other insect species is also common. Existing studies have shown that the differentiation of insect wing patterns is affected by many factors. It has been dete...
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IPC IPC(8): C12N15/10
Inventor 于居龙束兆林杨红福庄义庆姚克兵缪康赵来成
Owner ZHENJIANG AGRI SCI INST JIANGSU HILLY AREAS