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DNA (Deoxyribonucleic Acid) electrochemical biosensor and preparation method thereof

A biosensor, electrochemical technology, applied in the direction of electrochemical variables of materials, can solve the problems of high chemical stability, poor water solubility, difficult research and application of graphene, etc., to achieve the effect of good selectivity, low cost and high sensitivity

Inactive Publication Date: 2014-03-05
XINYANG NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is a strong van der Waals force between graphene sheets, which is prone to aggregation, and its chemical stability is high, and the surface is in an inert state, so its water solubility is poor, which has caused difficulties for further research and application of graphene.

Method used

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  • DNA (Deoxyribonucleic Acid) electrochemical biosensor and preparation method thereof
  • DNA (Deoxyribonucleic Acid) electrochemical biosensor and preparation method thereof
  • DNA (Deoxyribonucleic Acid) electrochemical biosensor and preparation method thereof

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preparation example Construction

[0029] The preparation method of described DNA electrochemical biosensor is:

[0030] (1) Gr-WS modified on the glassy carbon substrate of the biosensor 2 / Chitosan (CS) film;

[0031] First, the glassy carbon substrate was pretreated, and then 8 μL of Gr-WS 2 The / CS mixture was drop-coated on the glassy carbon substrate and dried in the air;

[0032] (2) DNA immobilization and hybridization:

[0033] Gr-WS 2 / CS film-modified glassy carbon substrate was soaked in gold colloid for 12 h, rinsed with water and dried, then soaked in DNA probe (ssDNA) solution at room temperature for 10 h, rinsed with buffer solution, soaked in mercaptoethanol Blocked in the solution for 2 h; take it out and rinse it with buffer solution. Then react the sensor with the target DNA at 30 °C for 50 min, wash with buffer solution, and dry to complete the immobilization and hybridization of DNA;

[0034] Gr-WS in the step (1) 2 The preparation method of / CS mixed solution is: 1 mg of Gr-WS 2 ...

Embodiment 1

[0038] Example 1: Preparation of DNA Electrochemical Biosensor

[0039] (1) Gr-WS modified on the glassy carbon substrate of the biosensor 2 / CS film

[0040] First, add CS to 1% (v%) acetic acid solution and stir for 1 h to prepare 1 mg / mL CS acetic acid solution. 1 mg of Gr-WS 2 The composite material was ultrasonically dispersed in 1 mL of double-distilled water for 30 min, and then 1 mL of the above-mentioned CS acetic acid solution was added and ultrasonically dispersed for 1 h to obtain Gr-WS 2 / CS mixture. The glassy carbon substrate was polished with polishing powder, then ultrasonically cleaned with ethanol and double distilled water, and dried in the air. Add 8 μL of the above Gr-WS 2 The / CS mixture was drop-coated on a glassy carbon substrate and air-dried.

[0041] (2) DNA immobilization and hybridization

[0042] Gr-WS 2 / CS film-modified glassy carbon substrate was soaked in gold colloid for 12 h, rinsed with water and dried, and then soaked in 1.0 × 10 ...

Embodiment 2

[0048] Such as Figure 5 As shown, the above-mentioned DNA electrochemical biosensor prepared by the present invention is used to measure different concentrations of target DNA. Determination conditions: The determination medium is 5.0 × 10 containing 0.1 mol / L KCl -3 mol / L of [Fe(CN) 6 ] 3? / 4? Solution; using differential pulse voltammetry (DPV), the measurement parameters are: pulse amplitude 0.05 V, pulse period 0.5 s, potential scanning range: -0.1~0.8 V. exist Figure 5 , the logarithm of the current intensity and the target DNA concentration is 1.0×10 -14 ~ 5.0×10 -10 Good linear relationship in the range of mol / L, the detection limit is 2.3 × 10 -15 mol / L. It shows that the biosensor can be used for sensitive detection of DNA sequence.

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Abstract

The invention relates to a DNA (Deoxyribonucleic Acid) electrochemical biosensor and a preparation method thereof. The preparation method comprises the following steps: (1) adding 1mg / mL Gr-WS2 composite material aqueous dispersion liquid into 1mg / mL chitosan acetic acid solution; carrying out ultrasonic dispersion for 1 hour; and taking 8 microliters of a mixed solution, dripping the solution on a pre-treated glassy carbon substrate and airing; and (2) immersing an electrode into a gold size for 12 hours; washing and airing; reacting the electrode with 1.0*10<-6>mol / L of a DNA probe at 25 DEG C for 10 hours; and washing and sealing by using 1.0*10<-3>mol / L of mercaptoethanol for 2 hours and then reacting with a target DNA at 30 DEG C for 50 minutes to prepare the DNA electrochemical biosensor. According to the DNA electrochemical biosensor provided by the invention, a Gr-WS2 nano composite material and gold nanoparticles are combined so as to be good for simultaneously expressing the advantages of the Gr-WS2 nano composite material and the gold nanoparticles; the DNA electrochemical biosensor has the advantages of good stability, high sensitivity, good selectivity, good repeatability, convenience for carrying, low cost and the like.

Description

technical field [0001] The invention relates to the field of electrochemical sensors, in particular to a DNA electrochemical biosensor and a preparation method thereof. Background technique [0002] DNA electrochemical biosensor is a kind of biosensor that uses DNA as the recognition element and recognizes through DNA sequence. It can convert the presence of target DNA into a detectable electrical signal. Compared with the traditional marker gene technology, the DNA electrochemical biosensor has the characteristics of faster, simpler, and pollution-free, and avoids the problems of inability to quantify and contamination leading to false positives. In addition, the DNA molecular recognition layer is very stable and easy to regenerate and synthesize, which makes DNA electrochemical biosensors play a vital role in the field of gene analysis and is widely used in medicine, food industry, environmental monitoring, drug research and development, etc. , has become a frontier topic...

Claims

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Application Information

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IPC IPC(8): G01N27/26G01N27/48
Inventor 刘彦明黄克靖刘玉洁王辉
Owner XINYANG NORMAL UNIVERSITY
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