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Preparation method for surface plasmon resonance DNA sensor based on graphene oxide

A surface plasmon and resonant sensor technology, applied in the direction of material excitation analysis, can solve the problems of low detection limit of single-stranded DNA, low fixed amount, and difficulty in meeting the requirements of ssDNA detection, and achieves good application prospects and detection methods. simple effect

Inactive Publication Date: 2013-11-27
JILIN UNIV
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Problems solved by technology

[0005] The main problems of the existing detection technology are: (1) In the SPR chip surface direct detection method, the immobilization amount of the probe ssDNA directly immobilized on the surface is small, and the detection limit for single-stranded DNA is not low enough
(2) The fluorescence quenching method uses fluorescent molecules to label probe ssDNA molecules, and the fluorescence quenching phenomenon occurs through the combination of fluorescent molecules and GO, so as to indirectly detect ssDNA molecules, which requires a complex fluorescent labeling process, special molecular biology equipment, and cost High, and cannot visually detect the concentration of ssDNA molecules
[0006] In fact, the development direction of sensing technology for detecting ssDNA is to improve sensitivity and specificity, reduce the detection limit, and simplify the detection process. However, due to the influence of sensing instruments and methods, it is impossible to use very simple sensing methods and sensor chips. Realize the ultra-low detection limit of ssDNA, it is difficult to meet the detection requirements of ssDNA in samples in scientific research and practical application

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  • Preparation method for surface plasmon resonance DNA sensor based on graphene oxide
  • Preparation method for surface plasmon resonance DNA sensor based on graphene oxide
  • Preparation method for surface plasmon resonance DNA sensor based on graphene oxide

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preparation example Construction

[0031] A method for preparing a graphene oxide-based surface plasmon resonance DNA sensor according to the present invention comprises the following steps:

[0032] First, the preparation of graphene oxide

[0033] The graphite oxide is dissolved in water, and a well-dispersed graphene oxide solution is obtained by ultrasonic stripping, and the concentration of the graphene oxide solution is 0.01-10 mg / ml.

[0034] Second, pass the prepared GO solution into the SPR sample cell, soak the SPR gold film chip in the GO solution for 1-16 hours, and realize the GO assembly on the surface of the SPR chip.

[0035] Third, nine different concentrations (10 -14 -10 -6 M) The base-matched target ssDNA solution is mixed with the competition reactant AuNPs-ssDNA solution, and the hybridization is incubated for 1-24 hours.

[0036] Fourth, pass the above nine solutions into the sample pool according to the high concentration to the low concentration, and the time for each concentration i...

Embodiment 1

[0040] Example 1: Detection method of target single-stranded DNA by SPR sensor assembled with GO

[0041]After GO was assembled on the surface of the SPR chip, when it interacted with single-stranded DNA solution and double-stranded DNA solution respectively, the amount of single-stranded DNA adsorbed on the GO surface was much larger than that of double-stranded DNA. Using ssDNA-functionalized gold nanoparticles (AuNPs-ssDNA) as a competitive reactant, the higher the concentration of the target single-stranded DNA mixed with it, the more double-stranded DNA will be formed, and the less the amount adsorbed to GO, the SPR resonance angle The smaller the change. When the target ssDNA was mismatched, the unmatched ssDNA and AuNPs-ssDNA were all adsorbed on the GO surface.

Embodiment 2

[0042] Embodiment 2: Preparation of gold nanoparticles

[0043] The preparation of 14nm gold nanoparticles is based on the traditional Frens-Turkevich method. In a 1L three-neck flask with a condenser, add 500mL of chloroauric acid with a concentration of 1mM, heat to boiling in an oil bath under condensing reflux and magnetic stirring, and rapidly Add 50 mL of sodium citrate with a concentration of 38.8 mM, the color changes from light yellow to purple, continue to stir and heat for 10 minutes, slowly cool to room temperature, and transfer to a jar for later use.

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Abstract

The invention discloses a preparation method for a surface plasmon resonance (SPR) DNA sensor based on graphene oxide, and belongs to the technical field of nanometer material biology. The technical problem to be solved in the preparation method is that the distinctive effect of graphene oxide-DNA is utilized, the surface plasmon resonance technology and the signal amplification mechanism of the AuNPs are applied, based on the competitive inhibition method, the phenomenon that the single-stranded DNA with different concentrations is absorbed on the surface of a sensing chip to cause SPR spectral change, and the single-stranded DNA is detected through the linear change of the resonance angle. Through the adoption of the preparation method, the SPR technology is adopted, the GO (Graphene Oxide) is utilized to assemble the surface of the chip, the single-stranded DNA is sensitively detected through adopting the competitive inhibition method and the AuNPs signal amplification effect, the concentration of the single-stranded DNA is quantitatively detected through analyzing the change of the SPR peak, and the detection limit is ultra-low. The preparation method has the advantages as follows: the instrument and equipment are low in cost, the cost is low, the operation is simple, the efficiency is high, the precision is high, and the detection limit is ultra-low.

Description

technical field [0001] The invention belongs to the technical field of surface plasmon resonance sensor single-stranded DNA biological detection, in particular to a graphene oxide SPR biosensor and a preparation method thereof. Background technique [0002] As the basic genetic material of organisms, DNA detection has great application value in biotechnology, environmental protection, life science and other fields. The research of DNA sensor has received extensive attention at home and abroad. Surface Plasmon Resonance (SPR) sensor (Surface Plasmon Resonance, SPR) is a resonance phenomenon produced by plasmon wave coupling irradiation light on the metal surface. The change of the dielectric constant and thickness of the metal film surface will affect the change of the SPR resonance curve. , so as to realize the sensing of the change of the medium environment on the metal surface. It has the characteristics of real-time, in-situ and label-free detection, fast detection, hig...

Claims

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Application Information

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IPC IPC(8): G01N21/63
Inventor 崔小强郑伟涛薛天宇
Owner JILIN UNIV
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