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Primer and probe for specific quantitative polymerase chain reaction (PCR) accurate detection of transgenic maize 98140 strain and method thereof

A technology for transgenic corn and detection methods, which is applied in biochemical equipment and methods, microbial measurement/inspection, DNA/RNA fragments, etc., can solve problems such as the lack of amplification detection transgenic corn 98140 accurate detection technology, and achieve informed protection The effect of right and option, high amplification efficiency and high accuracy

Inactive Publication Date: 2014-06-11
四川省农业科学院分析测试中心
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the detection technology of transgenic corn 98140 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 98140 and its products

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0035] The precise quantitative PCR detection method for transgenic maize 98140 strain mainly includes the following steps:

[0036] (1) Synthesize the following primers and fluorescent probes used in conjunction with the primers,

[0037] Upstream primer sequence, 98140event-F: 5'-GCGTTTTTTTGTGTGTGTATGTCTCT-3'

[0038] Downstream primer sequence, 98140event-R: 5'-CGTTTCCCGCCTTCAGTTTA-3'

[0039] Fluorescent probe sequence, 98140event-P: 5'-FAM-TGCTTGGTCTTTCTCTATCGATCCCCCTC-TAMRA-3'.

[0040] The synthesis concentrations of the primers and fluorescent probes in this example were both 10 μmol / l.

[0041] The nucleotide sequences of the above primers and fluorescent probes are designed for a specific site specific to the line of transgenic maize 98140 and its products, that is, the flanking site of the target gene and the recipient maize genome; through this design, the transgenic maize can be accurately detected The conversion event of 98140 in the middle.

[0042] (2) Prep...

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Abstract

The invention belongs to the technical field of an organism, relates to a quantitative analysis method of a gene, and particularly relates to a method for specific quantitative polymerase chain reaction (PCR) accurate detection of a transgenic maize 98140 strain. A PCR reaction system is prepared from a specific upstream primer sequence 98140event-F, a downstream primer sequence 98140event-R, a fluorescence probe sequence 98140event-P, a deoxyribonucleic acid (DNA) diluent of a 98140 strain, and TaqmanMastermix (2x) and water, and quantitative PCR detection is carried out. A Taqman quantitative PCR detection technique with high amplification efficiency and high accuracy is mainly built. The primer and the probe are applicable to supervision and inspection of domestic agricultural genetically modified organisms and products, inspection of genetically modified organisms and products of import and export ports, and biological ingredients detection of a transgenic gene 98140 strain in the internal imported 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 accurate detection method of specific quantitative PCR of maize 98140 line has become necessary. [0003] At present, the detection technology of transgenic corn 98140 mainly focuses on the common qualitat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q2561/101
Inventor 张富丽宋君雷绍荣郭灵安刘勇王东尹全刘文娟常丽娟
Owner 四川省农业科学院分析测试中心
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