Method and reagent for detecting transgenic maize strain VCO-01981-5

A technology of genetically modified corn and detection kits, applied in biochemical equipment and methods, microbe determination/inspection, DNA/RNA fragments, etc., can solve the problem of no strain-specific qualitative or quantitative detection methods, etc.

Active Publication Date: 2017-05-10
SHENZHEN CUSTOMS ANIMAL & PLANT INSPECTION & QUARANTINE TECH CENT
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AI Technical Summary

Problems solved by technology

This strain has not been approved by the Ministry of Agriculture of my country, and there is no corresponding strain-specific qualitative or quantitative detection method at home and abroad

Method used

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  • Method and reagent for detecting transgenic maize strain VCO-01981-5
  • Method and reagent for detecting transgenic maize strain VCO-01981-5
  • Method and reagent for detecting transgenic maize strain VCO-01981-5

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0033] 1. Materials and methods

[0034] 1. Test materials

[0035] In this example, DNA samples of transgenic maize lines VCO-01981-5, MIR162, MON89034, Bt176, Bt11, MON88017, MON810, MON863, T25, MIR604 and ES3272 were used for experiments. The above DNA samples were provided and preserved by the Animal and Plant Inspection and Quarantine Technology Center of Shenzhen Entry-Exit Inspection and Quarantine Bureau.

[0036] 2. Primer and Probe Design

[0037] In this example, specific detection primers and probes were designed based on the endogenous maize gene HMG and the specific sequence of the transgenic maize line VCO-01981-5, and the two probes were labeled with different fluorescent groups. Primers and probes were synthesized and modified by Sangon Bioengineering (Shanghai) Co., Ltd. The primers and probe sequences designed in this example are listed in Table 1.

[0038] Table 1 Endogenous genes and strain VCO-01981-5 specific primers and probes

[0039] n...

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Abstract

The invention discloses a method and a reagent for detecting the transgenic maize strain VCO-01981-5. The detection method provided by the invention comprises the following steps: adding mineral oil to a PCR reaction system, carrying out uniform mixing, transferring the mixture to a droplet generator for generating droplets, carrying out PCR reaction, after the reaction, reading an amplification signal of each droplet, and calculating the target DNA molecule number of a to-be-detected sample by adopting QuantaSoft software, wherein the PCR reaction system comprises strain VCO-01981-5 and HMG gene specific primers and probes, and the two probes are subjected to FAM and HEM fluorescence labeling. According to the quantitative determination method provided by the invention, the endogenous gene HMG and strain specific primers and probes matched for use are designed, by adopting a double-channel method, the molecule numbers of the endogenous gene HMG and the strain VCO-01981-5 in a maize gene group are quantified, the absolute content of the transgenic maize strain VCO-01981-5 is calculated, and thus the detection method has great significance for the research for the transgenic ingredients and the transgenic detection at the ports of china.

Description

technical field [0001] The application relates to the field of transgenic corn detection, in particular to a quantitative detection method and reagents for a transgenic corn strain VCO-01981-5. Background technique [0002] Since the commercialization of genetically modified crops in 1996, the acreage of genetically modified crops in the world has grown rapidly. As of 2013, the global area of ​​genetically modified crops has reached about 175 million hectares, an increase of 3% over 2012. According to the planting area statistics, about 81% of soybeans, 35% of corn, 30% of rapeseed and 81% of cotton in the world are genetically modified products. The top 6 countries with the planting area of ​​genetically modified crops are the United States, Brazil, Argentina, Canada, India and China. Along with the rapid development of genetically modified crops, the food safety issue of genetically modified crops has been a hot topic of discussion for many years, and directly affects in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6851C12Q2563/159C12Q2537/143C12Q2563/107
Inventor 凌杏园潘广叶奕优龙海章桂明程颖慧向才玉谢晓露
Owner SHENZHEN CUSTOMS ANIMAL & PLANT INSPECTION & QUARANTINE TECH CENT
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