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Preparation method and application of electrode for detecting transgenic CaMV35S promoter

A promoter and transgene technology, applied in measuring devices, material analysis through electromagnetic means, instruments, etc., can solve the problems of complex detection schemes, high detection costs, low sensitivity, etc., and reduce the probability of false negatives and false positives Effect

Inactive Publication Date: 2016-03-02
JIANGSU UNIV
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  • Summary
  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0009] The present invention aims at inventing a photoelectrochemical sensor integrating the advantages of label-free, high sensitivity, high selectivity, simple and easy operation, etc., and provides a quantitative detection containing The method of CaMV35S promoter transgenic crops and products solves the existing problems of high detection cost, complicated detection scheme, long detection time and low sensitivity

Method used

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  • Preparation method and application of electrode for detecting transgenic CaMV35S promoter
  • Preparation method and application of electrode for detecting transgenic CaMV35S promoter

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preparation example Construction

[0038] The preparation method of the graphene oxide used in the present invention is:

[0039] GO was prepared by the improved Hummers method: 1 g of natural flake graphite was added to 50 mL of concentrated H 2 S 2 o 4 (98%), cool to zero; slowly add 0.5gKNO 3 and 6gKMnO 4 . React for 4 hours under the condition that the reaction temperature is controlled not to exceed 10°C. Then the system was transferred to a constant temperature water bath at 35° C. for stirring for 2 h, 300 mL of deionized water was added, and the reaction was continued for 2 h at ≤80° C. With an excess of 5% H 2 o 2 Reduction of remaining KMnO 4 , and washed several times with 5% HCl, and finally fully washed with enough deionized water until the solution no longer contained SO 4 2- Ions (BaCl 2 No white precipitate was detected). The final product was transferred to a 65°C oven for drying and stored for future use.

[0040] The preparation method of rGO / Au is:

[0041] Weigh 25 mg of GO an...

Embodiment 1

[0043] SiO 2 Preparation of @CdTe

[0044] Take 1mg of 100nm silicon spheres, add them to 40mL of 0.4% diethylene glycol diacrylate (PDDA), stir for 1 hour, centrifuge at 7000 rpm for 5 minutes, wash with distilled water for 3 times, then take the precipitate and add it to 8 mL of distilled water containing 2.4 mg of CdTe was stirred and reacted for 1 hour, and the product was centrifuged, washed, and dried for later use.

Embodiment 2

[0046] Probe2 / SiO2 2 Preparation of @CdTe:

[0047] Take 1mL1mg / mL of SiO 2 @CdTe distilled water dispersion and 40 μL 0.2M N-hydroxysuccinimide (NHS) were added to 40 μL 0.4M ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), reaction 2 After 1 hour, centrifuge to remove excess EDC and NHS, add 100 μL of 10 μM Probe2tris-HCl buffer, and shake for 12 hours to obtain Probe2 / SiO 2 @CdTe.

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Abstract

The invention provides a preparation method and application of an electrode for detecting a transgenic CaMV35S promoter. The method includes the steps of: preparation of a reduced graphene oxide / gold nanocomposite RGO / Au; preparation of a silicon @ cadmium telluride nanocomposite Si@CdTe; preparation of a probe 1 (Probe 1) on an ITO electrode; modification of a target DNA (t-DNA) CaMV 35S promoter onto the probe 1 (Probe 1); and modification of a probe 2 (Probe2) onto the target DNA (t-DNA) CaMV 35S promoter. Through dual signal amplification at both ends of rGO / Au NPs and Si@CdTe, the sensitive detection of the transgenic CaMV35S promoter is realized; and in the concentration range of 0.05-100 pM, the concentration of CaMV35S and photocurrent present good linear relationship, and the detection limit can reach 0.017 pM.

Description

technical field [0001] The invention relates to the field of preparation of electrochemical electrodes, in particular to a preparation method and application of an electrode for detecting a transgenic CaMV35S promoter. Background technique [0002] Genetically Modified Crops (GMC for short) refers to the use of biotechnology to insert target genes identified and isolated from animals, plants or microorganisms into the plant genome to change its genetic composition and produce new agronomic traits. Since the first commercialization of genetically modified tomatoes in the United States in 1994, biotechnology has been widely used in modern agriculture. At present, there are more than 100 kinds of genetically modified organisms approved for commercial planting in the world, and 29 countries grow genetically modified crops. Many countries use some of the approved genetically modified crops as raw materials for food or feed production, and at the same time, the safety of genetica...

Claims

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Application Information

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IPC IPC(8): G01N27/327
Inventor 李雅琪王坤蔡建荣孙力
Owner JIANGSU UNIV
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