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Preparation method for synthesizing silver nanocluster electrochemical biosensor based on DNA signal amplification technique and application of electrochemical biosensor

A technology of biosensors and silver nanoclusters, which is applied in the field of preparation of electrochemical biosensors, can solve the problems of undisclosed electrochemical sensors, and achieve the effects of accelerating electron transfer, improving detection sensitivity, and high-sensitivity detection

Active Publication Date: 2017-04-19
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, there are no published reports on the research of electrochemical sensors based on graphene and DNA-AgNCs modified electrodes and the detection of hydrogen peroxide and TdT enzyme activity by electrochemical methods.

Method used

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  • Preparation method for synthesizing silver nanocluster electrochemical biosensor based on DNA signal amplification technique and application of electrochemical biosensor
  • Preparation method for synthesizing silver nanocluster electrochemical biosensor based on DNA signal amplification technique and application of electrochemical biosensor
  • Preparation method for synthesizing silver nanocluster electrochemical biosensor based on DNA signal amplification technique and application of electrochemical biosensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] A method for preparing an electrochemical biosensor based on DNA signal amplification technology to synthesize silver nanoclusters, the specific steps are as follows:

[0042] (1) Dispersion of graphene

[0043] Dissolve 8.0 mg of graphene in 8.0 mL of acetic acid buffer solution with a concentration of 0.2 M at pH=5.0~6.0, and ultrasonically disperse in an ultrasonic cleaner for 3.5 h to obtain a graphene dispersion;

[0044] (2) Preparation of silver nanoclusters (DNA-AgNCs)

[0045] a. Pipette 2.0 μL double distilled water, 8.0 μL DNA solution with a concentration of 12.0 μM, 1.5 μL dCTP solution with a concentration of 12.0mM, 1.5 μL 5×TdT reaction buffer and 0.3 μL TdT with a concentration of 15.0 U / μL Put the solution in a PCR tube, shake it for 3.5 min to mix the reagents evenly, then place it in a constant temperature water bath at 37 °C for 2.5 h, then place the PCR tube in a water bath at 75 °C for 15 min to terminate the reaction, and obtain the extended DNA...

Embodiment 2

[0051] A method for preparing an electrochemical biosensor based on DNA signal amplification technology to synthesize silver nanoclusters, the specific steps are as follows:

[0052] (1) Dispersion of graphene

[0053] Dissolve 5.0 mg of graphene in 10.0 mL of acetic acid buffer solution with a concentration of 0.1 M at pH=5.0~6.0, and ultrasonically disperse in an ultrasonic cleaner for 2 h to obtain a graphene dispersion;

[0054] (2) Preparation of silver nanoclusters (DNA-AgNCs)

[0055] a. Pipette 1.0 μL double-distilled water, 5.0 μL DNA solution with a concentration of 15.0 μM, 1.0 μL dCTP solution with a concentration of 15.0 mM, 1.0 μL 5×TdT reaction buffer and 0.4 μL TdT with a concentration of 10.0 U / μL Put the solution in a PCR tube, shake it for 2 minutes to mix the reagents evenly, then place it in a constant temperature water bath at 35°C for 3 hours, then place the PCR tube in a water bath at 70°C for 20 minutes to terminate the reaction, and obtain the extend...

Embodiment 3

[0061] A method for preparing an electrochemical biosensor based on DNA signal amplification technology to synthesize silver nanoclusters, the specific steps are as follows:

[0062] (1) Dispersion of graphene

[0063] Dissolve 10.0 mg of graphene in 5.0 mL of acetic acid buffer solution with a concentration of 0.3 M at pH=5.0~6.0, and ultrasonically disperse in an ultrasonic cleaner for 5 h to obtain a graphene dispersion;

[0064] (2) Preparation of silver nanoclusters (DNA-AgNCs)

[0065] a. Pipette 3.0 μL double distilled water, 10.0 μL DNA solution with a concentration of 15.0 μM, 1.0 μL dCTP solution with a concentration of 15.0 mM, 2.0 μL 5×TdT reaction buffer and 0.2 μL TdT with a concentration of 20.0 U / μL Put the solution in a PCR tube, shake it for 5 minutes to mix the reagents evenly, then place it in a constant temperature water bath at 38°C for 2 hours, then place the PCR tube in a water bath at 80°C for 10 minutes to terminate the reaction, and obtain the exten...

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Abstract

The invention discloses a preparation method for synthesizing a silver nanocluster electrochemical biosensor based on a DNA signal amplification technique and an application of the electrochemical biosensor, wherein the preparation method comprises the following specific steps: (1) dissolving graphene in an acetic acid buffer solution, and carrying out ultrasonic dispersion in an ultrasonic cleaner to obtain a graphene dispersion liquid; (2) dissolving a phosphoric acid buffer solution, a DNA reaction solution after extension and a silver nitrate aqueous solution in secondary distilled water, and mixing in a PCR tube evenly, then carrying out ice bath incubation to make silver ions combined with DNA, then adding a sodium borohydride aqueous solution, carrying out sustained oscillation to make the silver ions reduced, and carrying out light shielding reaction at room temperature to obtain silver nanoclusters; and (3) electrodepositing the graphene onto a bare glassy carbon electrode, then dropwise adding the DNA-AgNCs solution, and carrying out light shielding assembly and rinsing to obtain the product. The silver nanocluster electrochemical biosensor can be used for detection of the concentration of hydrogen peroxide and TdT enzymes, and has the advantages of good specificity, high sensitivity, high detection speed, accurate and reliable results and low cost.

Description

technical field [0001] The invention relates to an electrochemical sensor, in particular to a preparation method and application of an electrochemical biosensor based on DNA signal amplification technology to synthesize silver nano-clusters. Background technique [0002] Hydrogen peroxide is a common substance in nature, and it plays an important role in the fields of environment, industry, biotechnology food and clinical diagnostic analysis. Hydrogen peroxide itself has cytotoxicity, which will have a certain impact on biological processes such as activation of immune cells and cell apoptosis; many intermediate substances and final products that catalyze oxidoreductases are hydrogen peroxide; in industrial production, hydrogen peroxide The properties of oxidation, bleaching, disinfection and dechlorination make it widely used in the fields of textile, paper making, food and organic synthesis. Therefore, it is particularly important to develop a fast and efficient hydrogen ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/48B82Y30/00B82Y40/00
CPCB82Y30/00B82Y40/00G01N27/3278G01N27/48
Inventor 胡宇芳张青青郭智勇王邃杜春暖
Owner NINGBO UNIV
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