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Specific quantitative polymerase chain reaction (PCR) detection method for transgenic corn MON863 strain

A technology of MON863 and transgenic corn, which is applied in the fields of fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems that there is no precise detection technology of transgenic corn MON863, and achieve the protection of the right to know and the right to choose, high amplification efficiency and high accuracy Effect

Inactive Publication Date: 2012-04-11
四川省农业科学院分析测试中心
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Problems solved by technology

[0003] Moreover, the current detection technology for transgenic corn MON863 mainly focuses on ordinary qualitative PCR methods and standards, and there is no accurate quantitative PCR detection technology for detecting a specific site (gene sequence) specific to the genetically modified corn MON863 and its products.

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  • Specific quantitative polymerase chain reaction (PCR) detection method for transgenic corn MON863 strain

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Embodiment

[0029] The specific quantitative PCR detection method of transgenic maize MON863 line mainly comprises the following steps:

[0030] (1) Synthesize primers with the following nucleotide sequences and fluorescent probes used in conjunction with the primers,

[0031] Upstream primer sequence, Event863-F: 5'-CTACTTGTTCGGATGGGTGTTC-3'

[0032] Downstream primer sequence, Event863-R: 5'-CCTTTATCGCAATGATGGCA-3'

[0033] Fluorescent probe sequence, Event863-P: 5'-FAM-CCCCAAAGTGTACCAAGCTTTCCGAT-TAMRA-3'.

[0034] In this embodiment, the concentrations of the primers and the fluorescent probes after synthesis are both 10 μmol / l.

[0035] The nucleotide sequences of the above primers and fluorescent probes are designed for a specific site specific to the line of transgenic maize MON863 and its products, that is, the integration site of the exogenous gene and the maize genomic DNA. Through this design, the transgenic maize can be accurately detected Transformation events of MON863 in....

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Abstract

The invention belongs to a biological technology, and relates to a quantitative analytical method for genes, in particular to a specific quantitative polymerase chain reaction (PCR) detection method for a transgenic corn MON863 strain. The method comprises the following steps of: preparing a PCR reaction system from a specific forward primer Event863-F, a specific reverse primer Event863-R, a fluorescent probe Event863-P, deoxyribonucleic acid (DNA) diluent of the MON863 strain, TaqmanMastermix and water; and performing quantitative PCR detection. In the method, a Taqman quantitative PCR detection technology with high amplification efficiency and accuracy is established, and the method is suitable for the monitoring and inspection of transgenic organisms and transgenic products of domestic agriculture, the inspection of the transgenic organisms and the transgenic products of import and export ports and the detection of ingredients of organisms containing the transgenic MON863 strain in import raw materials of enterprises.

Description

technical field [0001] The invention belongs to biotechnology and relates to a quantitative analysis method of genes. Background technique [0002] Many countries in the world implement limited labeling and importation of genetically modified products, but there is no specific labeling threshold for genetically modified products in my country. In order to break the technical barriers to the trade of genetically modified products set up by the European Union and other countries and regions, at the same time make up for and improve the quantitative detection technology system of genetically modified organisms and products in my country, and better protect consumers' right to know and to choose genetically modified products, a new type of genetically modified Maize MON863 strain-specific quantitative PCR accurate detection technology has become necessary. [0003] Moreover, the current detection technology for transgenic maize MON863 mainly focuses on ordinary qualitative PCR m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/64
Inventor 宋君雷绍荣郭灵安尹全王东张富丽刘文娟常丽娟
Owner 四川省农业科学院分析测试中心
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