Method for noninvasive identification on genotype of insect

A genotype and insect technology, applied in the direction of biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as unsatisfactory, mechanical damage to insect bodies, inability to perform early identification of genotype identification accuracy, etc. Effects of mechanical damage

Pending Publication Date: 2020-09-11
HEBEI UNIVERSITY
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Problems solved by technology

[0005] The purpose of the present invention is to provide a method for non-invasive identification of insect genotypes to solve the problem that the existing identification methods will cause mechanical damage to the insect body, and the juvenile insect body will die due to mechanical damage, so early identification and genotype identification cannot be carried out. The problem of unsatisfactory accuracy

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  • Method for noninvasive identification on genotype of insect
  • Method for noninvasive identification on genotype of insect
  • Method for noninvasive identification on genotype of insect

Examples

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Embodiment Construction

[0021] In order to better illustrate the technical solution of the present invention, below, taking migratory locusts as an example, the novel insect genotype identification method of the present invention, that is, non-invasive, early and repeated genotype identification using molting is introduced in detail in conjunction with the accompanying drawings. Experimental operations and reagents not described in the examples were performed according to conventional operations in the field or the manufacturer's instructions.

[0022] The specific operation of the method for the non-invasive identification of migratory locust genotype of the present embodiment is as follows:

[0023] (1) Individuals with gene editing mutations:

[0024] Using CRISPR / Cas9 genome-directed editing technology to target migratory locust Orco Gene editing.

[0025] First, in vitro synthesis Orco Gene gRNA: Refer to the instructions of the gRNA synthesis kit (invitrogen, A29377), find a suitable targe...

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Abstract

The invention provides a method for noninvasive identification on a genotype of an insect. The method comprises the following steps: (a) separately feeding wild type insects and gene-edited insect mutants, and independently marking or feeding each insect individual; (b) employing that each ecdysis of each insect is regarded as once increase of age, and separately collecting ecdysis of corresponding insects from 1 age to an adult stage; (c) extracting genomic DNA from the ecdysis by using a DNA extraction kit; and (d) taking the genomic DNA extracted from the ecdysis as a template, designing primers according to a target gene sequence, carrying out PCR amplification, sequencing an amplification product, and then, identifying the genotype. According to the method, noninvasive, early-stage and repeated insect genotype identification can be achieved by using the ecdysis, and the problems that mechanical injury is caused by scissoring insect body tissue and insect bodies of a juvenile stagecannot be identified are solved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for noninvasive identification of insect genotypes. Background technique [0002] In recent years, with the continuous deepening of the research on the gene editing technology CRISPR / Cas9 system, the system has been widely used in many biological research fields due to its advantages of simple operation, time saving and high efficiency, especially for migratory locusts ( Locusta Migratoria ), Drosophila ( Drosophilidae ), silkworm ( Bombyx mori ), Aedes aegypti ( Aedes aegypti ) and other insect studies. After using the CRISPR / Cas9 genome-directed editing technology to edit the genome of insects, it is particularly important to identify the genotype of the edited individual. [0003] Currently, the sample tissues required for identification of insect mutants mainly use insect whole body or local tissues as materials to extract genomic DNA. For example, in the publi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6869
CPCC12Q1/6888C12Q1/6869C12Q2531/113
Inventor 魏丽亚康乐李静王彤欣周梦王笑笑
Owner HEBEI UNIVERSITY
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