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Probe sequence and kit for real time fluorescence PCR detection of transgenic rapeseed

A technology of real-time fluorescence and probe sequence, applied in the direction of microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problems of pollution, many operation steps, time-consuming and labor-intensive, etc., to avoid cross-contamination, accurate and sensitive results, avoid The effect of false positives

Inactive Publication Date: 2003-05-28
覃文 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The whole process of using this method to detect transgenic products is complicated, there are many steps, and post-PCR treatment is required, such as agarose gel electrophoresis and ethidium bromide staining, ultraviolet light observation results or detection by polyacrylamide gel electrophoresis or silver staining , not only need a variety of instruments, but also time-consuming and labor-intensive, the dye ethidium bromide used is harmful to the human body, and these complicated experimental processes provide opportunities for pollution and false positives, which seriously affect the correct judgment of the results

Method used

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  • Probe sequence and kit for real time fluorescence PCR detection of transgenic rapeseed
  • Probe sequence and kit for real time fluorescence PCR detection of transgenic rapeseed
  • Probe sequence and kit for real time fluorescence PCR detection of transgenic rapeseed

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Embodiment 1

[0019] A probe sequence and a kit for real-time fluorescent PCR detection of transgenic rapeseed to detect exogenous genes (transgenic components) of transgenic rapeseed.

[0020] Detection gene: Barnase gene.

[0021] Primers: 5'-ctgggtggcatcaaaagggaacc-3', 5'-tccggtctgaatttctgaagcctg-3'.

[0022] Probe: 5'-agacgtcgctccggggaaaa-3'.

[0023] Kit: a kit for detecting the Barnase gene of transgenic rapeseed made by using the probe.

Embodiment 2

[0025] A probe sequence and a kit for real-time fluorescent PCR detection of transgenic rapeseed to detect exogenous genes (transgenic components) of transgenic rapeseed.

[0026] Detected gene: modified CP4-EPSPS gene.

[0027] Primers: 5'-gacttgcgtgttcgttcttc-3', 5'-aacaccgttgagcttgagac-3'.

[0028] Probe: 5'-tgttccagaagaccgtgctc-3'.

[0029] Kit: a kit for detecting the modified CP4-EPSPS gene of transgenic rapeseed made by using the probe.

Embodiment 3

[0031] The probe sequence and kit for real-time fluorescent PCR detection of transgenic rapeseed detect the exogenous gene (transgenic component) of transgenic rapeseed.

[0032] Detection gene: FMV35S promoter.

[0033] Primers: 5'-aagcctcaacaaggtcag-3', 5'-ctgctcgatgttgacaag-3'.

[0034] Probe: 5'-ctccaaaccattagccaaaa-3'.

[0035] Kit: a kit for detecting the FMV35S promoter of the transgenic plant rapeseed made by using the probe.

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Abstract

A probe sequence and reagent kit for real-time fluorescent PCR detection of transgenic rapeseed is disclosed. The probe sequence for detecting Barnase gene, the modified CP4-EPSPS gene, the FMV355 promoter of repessed, and the endogenous gene of chloroplast, the maximal, minimal and preferable sequences, and the reagent kit are disclosed for quantitatively and qualitatively detecting the transgenic products. Its advantages are high sensitivity and correctness, high speed, and no cross infection.

Description

(1) Technical field: [0001] The invention relates to amplification and detection of genes. (two) background technology: [0002] With the continuous commercialization of genetically modified products, their safety issues have always been concerned by the world, and disputes continue to arise. In order to protect human health, eliminate consumer concerns, and facilitate international trade and commodity circulation, many countries, represented by the European Union, have formulated safety management regulations for such special products after a brief dispute, so that strict manage. Europe, Japan and many other countries have also formulated corresponding limit regulations. my country's Ministry of Agriculture has also issued management measures for genetically modified products this year. However, in order for consumers to understand or accept genetically modified products, various laws and regulations can be implemented, in addition to providing relevant information and i...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 覃文曹际娟
Owner 覃文
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