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Primer sequence and kit for real-time PCR detection of transgenic rape (seeds)

A primer sequence, transgenic technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of complex process of genetically modified products, affecting the correct judgment of results, and many operation steps, and achieve rapid and sensitive detection technology. Reliable results and the effect of avoiding cross-contamination

Inactive Publication Date: 2005-11-23
曹际娟 +2
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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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  • Primer sequence and kit for real-time PCR detection of transgenic rape (seeds)
  • Primer sequence and kit for real-time PCR detection of transgenic rape (seeds)

Examples

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Effect test

Embodiment 1

[0015] The primer sequences and kits used for SYBR® Green real-time PCR detection of transgenic rape (seed) RT73 strains are manufactured by conventional primer preparation and synthesis methods to detect exogenous structural genes (transgenic components) of transgenic rape (seed) RT73 strains.

[0016] Detected gene: exogenous structural gene of RT73 strain.

[0017] Primers: 5'-cca tat tga cca tca tact ca ttg ct-3', 5'-gct tat acg aag gca agaaaa gga-3'.

[0018] Kit: a kit for detecting exogenous structural genes of the transgenic rape (seed) RT73 line prepared by using the primer sequences and using conventional methods.

Embodiment 2

[0020] Primer sequences and kits for the SYBR® Green real-time PCR detection of transgenic rape (seed) MS8 strains are manufactured by conventional primer preparation and synthesis methods to detect exogenous structural genes (transgenic components) of transgenic rape (seed) MS8 strains.

[0021] Detected gene: exogenous structural gene of MS8 strain.

[0022] Primers: 5'-gac ctt tgc aat ttt ctc ttc agt act-3', 5'-gcc ttt tct tat cga cca tgtact c-3'.

[0023] Kit: a kit for detecting exogenous structural genes of transgenic rape (seed) MS8 line made by using the primer sequence.

Embodiment 3

[0025] The primer sequence and kit used for real-time PCR detection of SYBR(R)Green in rapeseed (seed) are manufactured according to conventional primer preparation and synthesis methods to detect endogenous genes in transgenic rapeseed (seed).

[0026] Detection gene: FatA gene.

[0027] Primers: 5'-ggt ctc tca gca agt ggg tga t-3', 5'-tcg tcc cga act tca tct gta a-3'.

[0028] Kit: a kit for detecting the endogenous FatA gene of rapeseed (seed) made by using the primer sequence.

[0029] Main instruments for detection: real-time PCR instrument, ultra-clean workbench, disinfection and sterilization pot, ice maker, nucleic acid and protein analyzer, high-speed refrigerated centrifuge, desktop small centrifuge, Mini personal centrifuge, low-temperature refrigerator, refrigerated refrigerator, Pure water device, double distilled water device, vortex shaker, micro sampler (0.5μL, 2μL, 10μL, 20μL, 100μL, 200μL, 1000μL), etc.

[0030] Main reagents for detection: 10×PCR buffer, M...

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Abstract

The present invention discloses the guide object sequence and reagent box used for real-time PCR inspection of transgenic rapeseed SYBR Green, providing the guide object sequence for the inspection of extraneous structure gene of transgenic rapeseed RT73 system, extraneous structure gene of MS8 system and endogenous gene of FatA, as well as the maxima, minima and optima selection sequence and the reagent box produced in accordance with guide object sequence. The present technology is used for qualitative and quantitative inspection of transgenic product and has the advantages of high resolution and avoiding cross pollution causing pseudo-positive reaction. SYBR Green real-time PCR technology employs complete closed-tube method, and PCR post treatment is not needed, avoiding cross pollution causing pseudo-positive reaction. SYBR Green dye is also very stable at the extreme temperature demanded by PCR reaction, which can efficiently distinguish specific product, nonspecific product and guide object dipolymer to compensate its bad specificity. The inspection technology is fast, sensitive, simple in operation, time saving and energy saving, accurate and reliable.

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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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 曹际娟刘善斌赵昕
Owner 曹际娟
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