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Method for researching helicoverpa armigera larva RNA interference efficiency

An RNA interference, cotton bollworm technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of difficult to eat completely, complicated operation, easy to damage larvae, etc., to improve the efficiency of RNA interference, stable Experimental results, strong operability effects

Inactive Publication Date: 2016-12-14
HUBEI UNIV
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AI Technical Summary

Problems solved by technology

Among them, the liquid drop feeding method uses a pipette to directly feed siRNA into the mouth of the cotton bollworm, which is complicated to operate and is easy to damage the larvae, and it is difficult to ensure complete feeding; the traditional feed mixing method mixes siRNA into the feed and carries out after a certain period of time. There is no quantification, and the amount of siRNA taken by each group of treatments or between each experiment cannot be controlled, and the amount of siRNA used is large; the surface smear feeding method applies siRNA on the surface of artificial feed, which has the disadvantages of slow action and low efficiency. In addition, the amount of siRNA introduced into the body of the cotton bollworm cannot be accurately introduced, the effect is not significant, and the repeatability of the experiment is poor, so it is difficult to obtain stable and accurate experimental results.

Method used

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  • Method for researching helicoverpa armigera larva RNA interference efficiency
  • Method for researching helicoverpa armigera larva RNA interference efficiency
  • Method for researching helicoverpa armigera larva RNA interference efficiency

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] This example is used to illustrate the effect of eating different amounts of ABCC2 siRNA on the ABCC2 gene of cotton bollworm

[0034] Experimental object: 3rd instar cotton bollworm larvae, 48 heads, repeated 3 times.

[0035] Dissolve ABCC2-siRNA and control Ncontrol siRNA in DEPC water respectively to make a solution with a concentration of 0.04-0.16 μg / μl, and use DEPC water as a control; prepare the artificial sample of cotton bollworm, and cut the feed into small pieces of 8-10 mg . Put the feed into the small well of the bioassay plate, and drop 2.5 μL of siRNA solution (or control NControl and DEPC water) on the feed. Select the 3rd instar larvae with uniform size and just after moulting, and place them in the above-mentioned small hole after starvation for 12 hours; after 6 hours, the cotton bollworm has completely eaten all the feed, repeat the above feeding; extract the cotton bolls after 12 hours Insect total RNA was reverse-transcribed into cDNA, and the ...

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Abstract

The invention relates to the field of biotechnology and particularly discloses a method for researching helicoverpa armigera larva RNA interference efficiency. The method for researching the helicoverpa armigera larva RNA interference efficiency is established by improving a method for feeding helicoverpa armigera larva siRNA (small interfering RNA), the lepidopteron helicoverpa armigera larva RNA interference efficiency is improved, and experimental results are stabilized. The method provided by the invention is efficient and simple, and is high in operability, the dosage of siRNA is definite, the repeatability is high, and technical supports for researching functions of genes of lepidopteron helicoverpa armigera are provided.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for studying RNA interference efficiency of cotton bollworm larvae. Background technique [0002] Cotton bollworm Helicoverpa armigera (Hübner) belongs to Lepidoptera Noctuidae insects, it is a worldwide distribution, polyphagous important agricultural pests, cotton bollworm damage cotton caused huge economic losses in the last century. The widespread application of transgenic Bt (Bacillus thuringiensist) cotton has effectively controlled the damage of cotton bollworm, but the resistance of cotton bollworm to Bt cotton is also increasing significantly, and at the same time, the harm of cotton bollworm to other non-transgenic crops is also increasing. intensified year by year. The use of gene interference technology to study the functional genes of cotton bollworm can provide a scientific basis for the control and management of cotton bollworm. [0003] siRNA (Small interfe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/158C12Q2600/166C12Q2600/178
Inventor 魏纪珍张江梁革梅
Owner HUBEI UNIV
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