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Transgenic maize multiple PCR screening detection primer and detection method

A technology of genetically modified corn and a detection method, which is applied in the field of multiple PCR screening and detection, can solve problems such as the lack of multiple PCR screening and detection strategies, and achieve the effects of low cost, high accuracy, and low false positive rate

Inactive Publication Date: 2015-03-11
四川省农业科学院分析测试中心
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Problems solved by technology

[0002] At present, the screening and detection technology for imported genetically modified corn and its products mainly focuses on the single-gene PCR method, and there is no research on multiple PCR screening and detection strategies and related screening and detection technologies for the detection of imported genetically modified corn and its products

Method used

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  • Transgenic maize multiple PCR screening detection primer and detection method
  • Transgenic maize multiple PCR screening detection primer and detection method
  • Transgenic maize multiple PCR screening detection primer and detection method

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Experimental program
Comparison scheme
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Embodiment

[0065] The multiplex PCR screening detection method for transgenic corn is as follows:

[0066] The double PCR screening method is as follows:

[0067] (a1) Synthesize the following primers:

[0068] P-CaMV 35S-F: 5'-GCTCCTACAAATGCCATCATTGC-3';

[0069] P-CaMV 35S-R: 5'-GATAGTGGGATTGTGCGTCATCCC-3';

[0070] T-NOS-F: 5'-TGAATCCTGTTGCCGGTCTT-3';

[0071] T-NOS-R: 5'-AAATGTATAATTGCGGGACTCTAATC-3';

[0072] (a2) preparing the DNA dilution of the corn to be tested;

[0073] (a3) Prepare PCR reaction system;

[0074] (a4) performing a PCR reaction;

[0075] (a5) The PCR reaction product is taken for capillary electrophoresis.

[0076] Among them, the total volume of the prepared PCR reaction system described in step (a3) ​​is 25 μL; specifically, it contains the following components: PCR Master Mix (2X), DNA diluent, each primer in step (a1), ddH 2 O; the amount of PCR Master Mix (2X) added was 12.5 μL, the amount of DNA diluent added was 2.0 μL, and the final concentration ...

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Abstract

The invention discloses a transgenic maize multiple PCR screening detection primer and detection method. The primer has relatively strong specificity, and can be used for multiple PCR screening detection analysis. The transgenic maize screening detection method which is simple and convenient to operate, good in extension performance and high in sensitivity is provided, and the transgenic maize multiple target detection is realized. The PCR amplification product is analyzed by utilizing capillary electrophoresis, the resolution ratio of the fragment size can reach a plurality of bases, and the resolution ratio is high. According to the primer and the detection method, a simple, convenient, effective, reliable and high-flux detection method is provided for the transgenic maize multiple target detection. A multiple PCR reaction system established by the experiment not only can detect foreign gene components of the imported transgenic maize, but also can be amplified to screening detection of other transgenic crops.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a multiplex PCR screening detection method capable of simultaneously detecting two and six exogenous genes. Background technique [0002] At present, the screening and detection technology for imported genetically modified corn and its products mainly focuses on the single-gene PCR method, and there is no research on multiple PCR screening and detection strategies and related screening and detection technologies for the detection of imported genetically modified corn and its products. Contents of the invention [0003] The purpose of the present invention is mainly to provide an accurate, fast and simple multiplex PCR rapid screening method for multiple target gene elements. [0004] The present invention realizes through following technical scheme: [0005] Multiplex PCR screening detection primers for transgenic maize, including the following primer sequences: [0006] Wherein, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2600/158C12Q2600/16C12Q2537/143
Inventor 尹全宋君刘勇雷绍荣郭灵安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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