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Primer, probe and method for specific quantitative polymerase chain reaction (PCR) accurate detection of transgenic maize DAS-40278-9 strain

A technology for transgenic corn and detection methods, applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problem that there is no amplification detection of transgenic corn DAS-40278-9 strain specificity, and achieve Protection of the right to know and right of choice, high amplification efficiency, and high accuracy

Inactive Publication Date: 2015-06-24
四川省农业科学院分析测试中心
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Problems solved by technology

[0003] At present, the detection technology of transgenic corn DAS-40278-9 mainly focuses on common qualitative PCR analysis methods and standards, and there is no specific site (gene sequence) for the amplification detection of transgenic corn DAS-40278-9 and its products. ) quantitative PCR precise detection technology

Method used

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  • Primer, probe and method for specific quantitative polymerase chain reaction (PCR) accurate detection of transgenic maize DAS-40278-9 strain
  • Primer, probe and method for specific quantitative polymerase chain reaction (PCR) accurate detection of transgenic maize DAS-40278-9 strain
  • Primer, probe and method for specific quantitative polymerase chain reaction (PCR) accurate detection of transgenic maize DAS-40278-9 strain

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Embodiment

[0029] The precise detection method of transgenic maize DAS-40278-9 strain-specific quantitative PCR comprises the following steps:

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

[0031] Upstream primer sequence, DAS-40278-9event-F: 5'-GTCCAGCACGAACCATTGAGT-3'

[0032] Downstream primer sequence, DAS-40278-9event-R: 5'-TTCATTGTATTCTGGCTTTGCTG-3'

[0033] Fluorescent probe sequence, DAS-40278-9event-P: 5'-FAM-CAACAGCACCGTACCTTGAAGCGGAAT-TAMRA-3'.

[0034] The synthesis concentrations of the primers and fluorescent probes in this example were 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 DAS-40278-9 and the product, i.e. the target gene and the flanking site of the recipient maize genome; Accurate detection of DAS-40278-9 transformation events in transgenic maize.

[0036] (2) Prepare the DNA di...

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Abstract

The invention belongs to the technical field of organisms, relates to a quantitative analysis method of a gene, and particularly relates to a specific quantitative polymerase chain reaction (PCR) accurate detection of a transgenic maize DAS-40278-9 strain. A PCR reaction system is prepared from deoxyribonucleic acid (DNA) diluents of designed upstream primer sequence DAS-40278-9event-F, a downstream primer sequence DAS-40278-9event-R, a fluorescent probe sequence DAS-40278-9event-P, a DAS-40278-9 strain, TaqmanMastermix(2x) and water, and quantitative PCR detection is carried out. A Taqman quantitative PCR detection technology with high amplification efficiency and high accuracy is mainly built, and the method is applicable to domestic agricultural transgenic organism and product supervision and inspection, transgenic organism and product inspection of inbound and outbound ports, and detection of transgenic DAS-40278-9 strain biotic components contained in inner-enterprise import raw materials.

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 precise detection method of maize DAS-40278-9 strain-specific quantitative PCR has become necessary. [0003] At present, the detection technology of transgenic corn DAS-40278-9 mainly focuses on common qu...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q2561/101
Inventor 尹全宋君雷绍荣郭灵安刘勇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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