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Specific quantitative PCR (Polymerase Chain Reaction) accurate detection method for transgenic corn 59122 strain

A technology of transgenic corn and detection method, which is applied in the biological field, can solve the problems of no amplification detection transgenic corn 59122 precision detection technology, and achieve the effects of protecting the right to know and the right to choose, high accuracy, and high amplification efficiency

Inactive Publication Date: 2013-08-28
四川省农业科学院分析测试中心
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AI Technical Summary

Problems solved by technology

[0003] At present, the detection technology of transgenic corn 59122 mainly focuses on common qualitative PCR analysis methods and standards, and there is no accurate quantitative PCR detection technology for amplifying and detecting a specific site (gene sequence) specific to the strain of transgenic corn 59122 and its products

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0029] The precise quantitative PCR detection method for transgenic maize 59122 strain mainly includes 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, 59122event-F: 5'-AATTCGCCCTATAGTGAGTCGTA-3'

[0032] Downstream primer sequence, 59122event-R: 5'-GATAAACAAACGGGACCATAGAA-3'

[0033] Fluorescent probe sequence, 59122event-P: 5'-FAM-CGCAATTCAGTACATTAAAAACGTCCGC-TAMRA-3'.

[0034] The synthesis concentrations of the primers and fluorescent probes in this example 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 corn 59122 and its products, that is, the integration site of the exogenous gene and the corn genomic DNA. Through this design, the transgenic corn can be accurately detected Transformation events in 59122.

[0036]...

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Abstract

The invention belongs to a biological technology, and relates to the quantitative analyzing method of genes, in particular to a specific quantitative PCR (Polymerase Chain Reaction) accurate detection method for a transgenic corn 59122 strain. In the method, a PCR reaction system is prepared by adopting a designed specific upstream primer 59122event-F, a downstream primer 59122event-R, a fluorescent probe 59122event-P, a DNA diluent of the 59122 strain, TaqmanMastermix and water for performing quantitative PCR detection. In the method, a Taqman quantitative PCR detection technology with high amplification efficiency and high accuracy is mainly established; and the method is suitable for supervision and inspection of domestic agricultural transgenic organisms and products, inspection of entry-exit port transgenic organisms and products, and biological component detection of enterprise internal import raw materials containing transgenic 59122 strains.

Description

technical field [0001] The invention belongs to the field of 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 The accurate detection method of maize 59122 strain-specific quantitative PCR has become necessary. [0003] At present, the detection technology of genetically modified corn 59122 mainly focuses on common qua...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 宋君王东雷绍荣郭灵安刘勇尹全刘文娟张富丽常丽娟
Owner 四川省农业科学院分析测试中心
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