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A rapid detection kit for simultaneous detection of multiple transgenic rapeseed lines and its application

A kit and transgenic technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of high detection cost, unsuitable multi-strain screening and detection of genetically modified products, environmental pollution, etc. Achieving good stability and suitable for commercial promotion

Active Publication Date: 2021-03-02
DALIAN NATIONALITIES UNIVERSITY +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 rapid detection kit for simultaneous detection of multiple transgenic rapeseed lines and its application
  • A rapid detection kit for simultaneous detection of multiple transgenic rapeseed lines and its application
  • A rapid detection kit for simultaneous detection of multiple transgenic rapeseed lines and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: PCR detection method and sensitivity, specificity experiment

[0033] (1) Materials and methods

[0034]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) ), transgenic rapeseed Ms1 (AOCS0711-A3), genetically modified rapeseed Ms8 (AOCS0306-F7), genetically modified rapeseed Rf3 (AOCS0306-G6), transgenic rapeseed T45 (AOCS0208-A6), genetically modified rapeseed Rf1 (AOCS0711-B2), transgenic rapeseed GT73 (AOCS0304-B2), transgenic rape MON88302 (AOCS1011-A), non-transgenic rape (AOCS 0304-A2), transgenic soybean MON89788 (AOCS 0906-B2), non-transgenic soybean (AOCS 0906-A), transgenic corn GA21 ( AOCS0407-B), non-transgenic maize (AOCS0411-CD) and transgenic rapeseed 73496 (ERM-BF434e) purchased from European Reference Materials (ERM...

Embodiment 2

[0054] Example 2: Verification of microfluidic chip stability

[0055] 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.

[0056] The detection chips for CRUA, transgenic rapeseed RF2, and transgenic rapeseed T45 were placed at room temperature (25°C) and -20°C for 3 months, 6 months, and 12 months, respectively, to test the stability of the kit. The verification experiment was carried out with transgenic rapeseed Topas19 / 2 (AOCS0711-D4) at a concentration of 5ng / uL. 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...

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Abstract

The invention relates to a PCR micro-fluidic chip special for identification of transgenic rapeseed strains. According to the PCR micro-fluidic chip special for identification of the transgenic rapeseed strains, a series of common transgenic rapeseed element PCR amplification primers, with high specificity and high accuracy, suitable for the micro-fluidic chip are obtained through designing and screening, and through freeze-drying, the primers and the PCR micro-fluidic chip are combined, so that high-throughput screening detection of an endogenous gene and multiple exogenous genes is achievedin a primary PCR amplification reaction. The provided PCR micro-fluidic chip with the pre-cryopreserved primers has good stability in both a normal temperature situation and a situation of repeated freezing and thawing, and has good commercial application value.

Description

technical field [0001] The invention relates to the technical field of agricultural transgene detection, in particular to a microfluidic fluorescent PCR chip kit containing a primer set capable of simultaneously screening 10 transgenic rapeseed strains and an application thereof. Background technique [0002] 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 modified crops to the environment, society and economy have been questioned in many ways. Therefore, it has become an international practice to strengthen the safety supervision of genetically modified produ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2600/16C12Q2565/629C12Q2537/143C12Q2563/107
Inventor 曹际娟徐君怡郑秋月朴永哲杨莉莉
Owner DALIAN NATIONALITIES UNIVERSITY
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