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Transgenic product surface plasma resonance biosensor detecting method

A surface plasmon and biosensor technology, applied in the field of detection of genetically modified products, can solve the problems of human and environmental hazards, lack of specificity and sensitivity, etc., and achieve the effects of continuous analysis, shortened detection time, and fast and accurate process.

Inactive Publication Date: 2005-02-16
XIAMEN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the detection of polygenic PCR products still uses the traditional gel electrophoresis analysis method, which is simple and easy to implement, but lacks specificity and sensitivity, and is likely to cause harm to humans and the environment

Method used

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  • Transgenic product surface plasma resonance biosensor detecting method
  • Transgenic product surface plasma resonance biosensor detecting method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Selection of appropriate primers and probes is the key to successful DNA analysis and detection. For the most commonly used exogenous DNA in transgenic products, such as CaMV 35S promoter and Tnos terminator sequence, through repeated combination and adjustment of various parameters, through the software Primer Express TM Search and analyze with OLIGO 4.0, and compare each oligonucleotide, and select primers and probes that meet the requirements, see Tables 1 and 2 for details.

[0025] Primer number

target DNA

Primer sequence (5'-3')

35S-1 / 2

CaMV 35S promoter

GCT CCT ACA AAT GCC ATC A

GAT AGT GGG ATT GTG CGT CA

nos-1 / 2

Tnos terminator

GAA TCC TGT TGC CGG TCT TG

TTA TCC TAG TTT GCG CGC TA

[0026] Probe No.

target DNA

Probe sequence (5'-3')

35S-P1

CaMV 35S promoter

GAG GAG CAT CGT GGA AAA AGA AG

nos-P1

Tnos terminator

GCA TGA CGT TAT TT...

Embodiment 2

[0030] Design the terminal biotinylated specific probe 35S-P1 and immobilize it on the surface of the chip, add the CaMV 35S-PCR amplification products of plasmids pBI121(S-121) and pUC19(S-19) respectively, and monitor on-site by BIAcore X instrument The optical signal change process generated by the hybridization of the probe and the amplification product of the plasmid. The results show that the 35S-P1 probe can specifically hybridize with the amplification product of S-121 and produce an obvious hybridization curve, but has no obvious hybridization signal with the amplification product of S-19 (see figure 1 ).

[0031] SPR technology suggests that by detecting the complementary binding between DNA molecules, that is, detecting the hybridization reaction process between PCR products and probes, the specific detection of PCR products can be realized; online and real-time monitoring of the hybridization reaction process by instruments can realize Automated analysis of PCR pr...

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Abstract

The detecting method of transferring gene product's surface plasma resonate biological sensor, which relates to a detecting method adopts to transferring gene product with surface plasma resonate biological sensor. Its procedure is designing and preparing ssDNA probe modified by tip biotin;using absorption to fix to probe on the surface of chip and forming monomolecular layer to execute detecting chip; adding sampling DNA PCR product to monitor cross reaction and observe analytic result after elution. Leading to the detecting of transferring product,the process is shortcut, accurate and real time response. Not only the psitive result of the determined PCR detected can be verificated, but also the selecting experiment can use the transferring geng product's specificity detecting probe. As compared to general gene detection, the probe is immobilizing, uses repeatedly and is low cost ;the sample needn't deal with in advance and extra reagent, can analyse directly; answering quickly, cutting detecting time and analyzing continuously to make true on line and real time monitoring of DNA; the analysis work is simple and is easy to automatic measurement.

Description

technical field [0001] The invention relates to a detection method of a genetically modified product, in particular to a detection method using a surface plasmon resonance biosensor for the genetically modified product. Background technique [0002] With the rapid development of genetically modified crops, the environmental safety and food safety of genetically modified products have attracted widespread attention from governments and the public (Anklam E, Gadani F, Heinze P, et al, Analytical methods for detection and determination of genetically modified organisms in agricultural crops and plant-derived food products. Eur. Food Res. Technol., 2002, 214, 3-26) In order to protect consumers' right to know and right to choose, to ensure the safety of people's dining tables, and to ensure the healthy development of international trade in agricultural products, countries around the world Both have increased research investment in the detection and safety evaluation of genetically...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 苏文金刘光明宋思杨
Owner XIAMEN UNIV
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