Method for detecting paddy rice transgenic ingredients

A technology of genetically modified components and detection methods, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve problems such as failure, wrong administrative penalties for non-transgenic varieties, and achieve accurate and reliable results

Inactive Publication Date: 2016-05-18
JIANGHAN UNIVERSITY
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Problems solved by technology

[0005] There are many kinds of genetically modified ingredients, and the existing technology can only detect one or a few of them at a time. Therefore, it is necessary to test the possible multiple genetically modified ingredients one by one before they can be determined as "non-genetically modified" products; The transformed genetically modified components are not the exogenous gene itself. Therefore, for unmarked transgenes, the existing

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  • Method for detecting paddy rice transgenic ingredients
  • Method for detecting paddy rice transgenic ingredients
  • Method for detecting paddy rice transgenic ingredients

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Embodiment

[0029] The Biolaphosresistance (Bar) gene was introduced into Nipponbare rice varieties by the Agrobacterium-mediated method, and after selfing and purification for 3 generations, modified Nipponbare rice varieties seeds were obtained as the plant varieties to be tested in this example. Among them, Nipponbare rice is a standard transgenic rice recipient variety, and almost every transgenic laboratory has Nipponbare rice seeds. The Nipponbare rice seeds used in this example are preserved and propagated by Jianghan University.

[0030] The specific steps for detecting genetically modified components of rice varieties are as follows:

[0031] Step 1. Determine the genetically modified components that need to be detected in the test sample, the endogenous standard gene in the test sample, and the exogenous standard gene of the test sample. The specific method is as follows: the test sample is the root, stem, leaf, and At least one of seeds and flowers. Specifically, the sample to be te...

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Abstract

The invention discloses a method for detecting paddy rice transgenic ingredients, and belongs to the field of biotechnology. The method comprises: determining a transgenic ingredient, an endogenous standard gene and an exogenous standard gene; preparing a multiple-amplification primer; carrying out sampling and mixing to obtain a mixed sample; extracting a genome of the mixed sample, and adding the exogenous standard gene to obtain mixed nucleic acid; constructing a high-flux sequencing library; carrying out high-flux sequencing on the high-flux sequencing library to obtain a sequencing fragment group; obtaining the number of sequencing fragments of the transgenic ingredient, the number of sequencing fragments of the endogenous standard gene and the number of sequencing fragments of the exogenous standard gene in a to-be-tested sample; according to the number of the sequencing fragments of the exogenous standard gene, determining whether an experiment is successful or not, and calculating the content of the transgenic ingredient in the to-be-tested sample; and according to the content of the transgenic ingredient, determining whether the to-be-tested sample contains the transgenic ingredient. According to the method, various transgenic ingredients in the to-be-tested sample can be quantitatively detected at the same time.

Description

Technical field [0001] The invention relates to the field of biological detection, in particular to a detection method of rice transgenic components. Background technique [0002] Rice is the main food crop in my country, and the genetically modified technology of rice and its products has matured. The planting and promotion of genetically modified rice varieties require strict approval. Therefore, genetically modified rice needs to be supervised, and genetically modified component testing is the technical support for genetically modified supervision. [0003] The existing genetically modified component detection technology mainly detects nucleic acid or protein. Among them, the main process of nucleic acid-based detection methods is: extract nucleic acid from the sample to be tested, and use PCR (Polymerase Chain Reaction, polymerase chain reaction) method to amplify the transgene in the sample Ingredients, using real-time quantification, agarose electrophoresis or chip technology...

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6869C12Q2545/101C12Q2545/113C12Q2537/143
Inventor 方治伟彭海周俊飞
Owner JIANGHAN UNIVERSITY
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