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Novel detection kit for transgenic aspergillus oryzae and detection method thereof

A technology of Aspergillus oryzae and transgene, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc. It can solve the problems of EB dye pollution, time-consuming, and many steps to avoid pollution and detect Intuitive results

Inactive Publication Date: 2014-01-08
ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

However, the amplified product needs gel electrophoresis and ultraviolet gel imaging, which requires many steps, takes a long time, and there is the problem of EB stain contamination.

Method used

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  • Novel detection kit for transgenic aspergillus oryzae and detection method thereof
  • Novel detection kit for transgenic aspergillus oryzae and detection method thereof
  • Novel detection kit for transgenic aspergillus oryzae and detection method thereof

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Experimental program
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Effect test

Embodiment 1

[0043] Embodiment 1, the detection of transgenic Aspergillus oryzae

[0044] DNA extraction: Qiagen DNeasy plant mini kit was used to extract fungal genomic DNA according to the steps, dissolved in 100uL TE, and stored at -20°C for later use.

[0045] PCR amplification: respectively with ddH 2 O, A.ory1, A.ory2, A.ory3, A.ory4, A.ory5, A.ory6 and NZ-A.ory genomic DNA were used as templates for PCR amplification.

[0046] PCR reaction system:

[0047]

[0048] PCR reaction conditions: pre-denaturation at 94°C for 1 min, denaturation at 94°C for 30 s, annealing at 56°C for 30 s, extension at 72°C for 30 s, 30 cycles, and extension at 72°C for 5 min.

[0049] Probe hybridization: After amplification, add 1 μl of probe (10 μmol / L), denature at 94°C for 30 seconds, and anneal at 55°C for 30 seconds. Test strip test: Take 4 μL of amplification and hybridization products and add them to the sample pad of the test strip, then add 2-3 drops of buffer solution, and read and record...

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Abstract

The invention relates to a novel detection kit for transgenic aspergillus oryzae and a detection method thereof, and relates to the technical field of transgenic microbiological detection. By designing a labeled primer and a probe for specific sequences of transgenic aspergillus oryzae and combining PCR amplification with nucleic acid test strip detection technology, the invention realizes accurate, high-efficient and rapid detection of transgenic aspergillus oryzae. The method has strong specificity, high sensitivity and good stability, and plays an important role in improving accuracy and shortening detection time for transgenic aspergillus oryzae detection. The detection kit is convenient and rapid for use, simple in principle and operation, and applicable to rapid transgenic aspergillus oryzae detection.

Description

technical field [0001] The invention relates to the technical field of transgenic microorganism detection, and provides a kit for rapidly detecting transgenic aspergillus oryzae by combining PCR amplification and nucleic acid test strip technology, which is suitable for rapid detection of transgenic aspergillus oryzae. Background technique [0002] Aspergillus oryzae (Aspergillus oryzae), belonging to the subphylum Deuteromycota, Hyphospora, Hyphospora, Polygonaceae, Aspergillus genus, is a common species in fungi. Widely distributed, mainly in food, fermented food, spoilage organic matter and soil. It is the production strain of my country's traditional brewed food sauce and soy sauce. It can also produce amylase, protease, pectinase and kojic acid, etc. It is a very important industrial strain. [0003] At present, the detection technology of genetically modified microorganisms in enzyme preparation products still mainly relies on the traditional morphological detection ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
CPCC12Q1/686C12Q2565/625C12Q2531/113
Inventor 张裕君赵卫东刘国红贺艳郑文杰
Owner ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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