Primers, probes and methods for accurate detection of specific quantitative PCR of transgenic maize mon88017 strain

A technology of genetically modified corn and detection methods, which is applied in the directions of biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. To achieve the effect of protecting the right to know and the right to choose, high amplification efficiency and high accuracy

Inactive Publication Date: 2018-07-24
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 MON88017 mainly focuses on the common qualitative PCR analysis method, and there is no high-sensitivity quantitative PCR accurate detection technology for amplifying and detecting a specific site (gene sequence) specific to the line of transgenic corn MON88017 and its products.

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  • Primers, probes and methods for accurate detection of specific quantitative PCR of transgenic maize mon88017 strain
  • Primers, probes and methods for accurate detection of specific quantitative PCR of transgenic maize mon88017 strain
  • Primers, probes and methods for accurate detection of specific quantitative PCR of transgenic maize mon88017 strain

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Experimental program
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Embodiment

[0035] The method for accurate detection of transgenic maize MON88017 strain-specific quantitative PCR mainly includes the following steps:

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

[0037] Upstream primer sequence: Event MON88017-Forward: 5'-CGCTAGCAGCTCTCCTCAA-3';

[0038] Downstream primer sequence: Event MON88017-Reverse: 5'-CCGGACATGAAGCCATTTACA-3';

[0039] Fluorescent Probe Sequence:

[0040] Event MON88017-Probe: 5'-FAM-CTTTTTTGCCGGAGTATGACGGTGACG-3'.

[0041] The synthesis concentrations of the primers and fluorescent probes in this example were both 10 μmol / l.

[0042] The nucleotide sequences of the above primers and fluorescent probes are designed for a specific site specific to the line of transgenic maize MON88017 and its products, that is, the target gene and the flanking site of the recipient maize genome; through this design, the transgenic maize can be accurately detected The transformation...

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Abstract

The invention belongs to the field of biotechnology and relates to a method for quantitative analysis of genes, in particular to a precise detection method for specific quantitative PCR of transgenic maize MON88017 strain. The invention adopts the designed specific upstream primer sequence Event MON88017-Forward, downstream primer sequence Event MON88017-Reverse, fluorescent probe sequence Event MON88017-Probe, DNA dilution of MON88017 strain and Taqman Master mix (2×) and water to prepare PCR reaction system for quantitative PCR detection. The present invention mainly establishes a Taqman quantitative PCR detection technology with high amplification efficiency and high accuracy, which is suitable for the supervision and inspection of domestic agricultural transgenic organisms and products, the inspection of genetically modified organisms and products at entry and exit ports, and the imported raw materials containing transgenic MON88017 strains in enterprises. Biological component testing.

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 MON88017 strain-specific quantitative PCR has become necessary. [0003] At present, the detection technology of transgenic maize MON88017 mainly focuses on the ordinary ...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/6895
Inventor 王东宋君雷绍荣郭灵安刘勇常丽娟张富丽尹全刘文娟
Owner 四川省农业科学院分析测试中心
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