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A kit for rapid screening of genetically modified ingredients in feed and its application

A kit and gene technology, applied in the field of microfluidic real-time fluorescent PCR chip kit, can solve the problems of multi-line screening and detection of unsuitable genetically modified products, high detection cost, environmental pollution, etc. The effect of stability

Active Publication Date: 2021-03-05
大连海关技术中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Ordinary PCR method has been gradually replaced by real-time fluorescent PCR detection method due to its shortcomings such as cumbersome operation, polluted environment, and low sensitivity.
However, the detection cost of single-channel TaqMan real-time fluorescent PCR method is high, and the steps are cumbersome, and it is not suitable for multi-line screening detection of genetically modified products.

Method used

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  • A kit for rapid screening of genetically modified ingredients in feed and its application
  • A kit for rapid screening of genetically modified ingredients in feed and its application
  • A kit for rapid screening of genetically modified ingredients in feed and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: PCR detection method and sensitivity, specificity experiment

[0031] (1) Materials and methods

[0032]Transgenic standard products were purchased from American Standard Materials AOCS or European Standard Materials ERM, including: transgenic rapeseed Topas19 / 2 (AOCS0711-D4), transgenic rapeseed RF2 (AOCS0711-C3) purchased from American Oil Chemists' Society (AOCS, Urbana, USA) ), genetically modified soybean A5547-127 (AOCS 0707-C3), genetically modified soybean A2704-12 (AOCS 0707-B11), genetically modified soybean MON87701 (AOCS 0809-A), genetically modified soybean CV127 (AOCS0911-D), genetically modified soybean MON87769 (AOCS 0809-B), transgenic soybean FG72 (AOCS 0610-A4), non-transgenic rapeseed (AOCS 0304-A2), non-transgenic soybean (AOCS 0906-A), non-transgenic corn (AOCS0411-CD) and other genetically modified reference materials; purchased from European Reference Materials (ERM) transgenic corn 59122 (ERM-BF424d), transgenic soybean DP356043 (...

Embodiment 2

[0053] Example 2: Verification of microfluidic chip stability

[0054] The stability of the microfluidic chip of the present invention is verified, and the microfluidic chip that is not added with taurine and freeze-dried is used as a control to verify the conditions of the chip at room temperature (25°C), -20°C and repeated freezing and thawing Under the chip detection performance.

[0055] Using 0.1% concentration of transgenic soybean DP305423 (ERM BF426d, 10% concentration) sample and the detection chip for detecting 18sRNA endogenous gene and DP305423 3' border sequence were used for experiments to test the stability of the kit. The experimental results are shown in Table 3 below. When the microfluidic chip of the present invention is placed at room temperature (25°C) for 3 months, 6 months, and 12 months, there is no significant difference in the Ct value and Tm value, and the detection sensitivity is also not obvious. difference; when stored at -20°C for 3 months, 6 mo...

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Abstract

The invention relates to a PCR microfluidic chip specially used for identification of genetically modified components in feed. The invention designs and screens a series of high-specificity and high-accuracy PCR amplification primers for common transgenic components suitable for microfluidic chips, and combines the primers with PCR microfluidic chips by freeze-drying to achieve a single PCR High-throughput screening detection of endogenous genes and multiple exogenous genes in amplification reactions. The PCR microfluidic chip with pre-frozen primers provided by this application has good stability at room temperature and repeated freezing and thawing, and has good commercial application value.

Description

technical field [0001] The invention relates to the technical field of transgene detection, in particular to a microfluidic real-time fluorescent PCR chip kit for simultaneously screening 10 kinds of transgenic elements in feed and related applications thereof. Background technique [0002] Feed is an important material guarantee for animal food, and the quality and safety of feed directly affects the safety of animal food. At present, many feed products use genetically modified crops as raw materials. [0003] Genetically modified crops began to be commercially planted on a large scale in 1996, and the global planting area reached 189.8 million hectares in 2017. GMO crops have been accompanied by mixed voices since they were first commercialized. On the one hand, genetically modified crops have indeed greatly increased the yield of crops, reduced the use of pesticides, and protected the environment; but on the other hand, the potential uncertainties caused by genetically ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895
Inventor 徐君怡曹际娟郑秋月朴永哲汪霖
Owner 大连海关技术中心
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