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DNA electrochemical sensor and preparation method thereof

An electrochemical and sensor technology, applied in the field of deoxyribonucleic acid electrochemical sensor and its preparation, can solve the problems of inability to realize SNP detection and insufficient detection sensitivity

Inactive Publication Date: 2007-11-28
JIANGSU WUZHONG HIGH & NEW TECH IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection sensitivity of this sensor can reach 10 pmol / L, but it can only be applied to the target DNA to be tested with sulfhydryl groups, or it is modified with sulfhydryl groups before detection, so the practical application has certain limitations; Sensitive method, its detection sensitivity is still very insufficient, and it is not yet possible to realize SNP detection

Method used

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  • DNA electrochemical sensor and preparation method thereof
  • DNA electrochemical sensor and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] As shown in Figure 1, steps A, B, and C are used to prepare the DNA electrochemical sensor of the present invention.

[0039] (1) Electrode pretreatment and assembly of capture probe DNA

[0040] Firstly, the gold electrode was polished with alumina abrasive suspension on a grinding cloth, then ultrasonically cleaned in ethanol and MilliQ water respectively, and finally the electrode was cleaned electrochemically to remove residual impurities. After blowing dry with nitrogen, add 0.2 μM thiol-modified oligonucleotide probe assembly solution [pH7.2 phosphate buffer (PBS), where the NaCl concentration is 0.1M] dropwise on the electrode, and let it stand at room temperature for about 60 Minutes, and then soak in 2mM mercaptohexanol aqueous solution for about 2 hours, take it out and rinse it with MilliQ water.

[0041] (2) hybridization

[0042] First mix the DNA of the target sequence with the AuNP-DNA solution (AuNP diameter is 20 nanometers), pre-hybridize at 25°C for...

Embodiment 2

[0050] (1) Assembly of capture probe DNA on gold electrode surface

[0051] A PBS solution of sulfhydryl-modified capture probes at pH 7.4 was added dropwise on a clean and dry gold electrode, and assembled at room temperature for 60 minutes. Among them, by changing the concentration of the capture probe in the assembly solution to 0.2, 2.0, 5.0 μM and the concentration of NaCl to 0.1M, 1.0M, 1.0M respectively, DNA modified electrodes with different densities were assembled, divided into high (1.2×10 13 molecule / cm 2 ), Medium (6.0×10 12 molecule / cm 2 ), low (1.2×10 12 molecule / cm 2 ) three densities (as shown in Figure 2 C), and then soaked in 1mM mercaptohexanol aqueous solution for 3 hours, rinsed with MilliQ water after taking it out.

[0052] (2) hybridization

[0053] Mix target sequence DNA with AuNP-DNA solution (AuNP diameter is 20 nm), and pre-hybridize at 37° C. for 30 minutes. The rest is the same as in Example 1.

[0054] (3) Electrochemical detection

[0...

Embodiment 3

[0060] The pH of the capture probe solution is 7.8, assembled for 30 minutes, and then soaked in 1mM mercaptohexanol aqueous solution for 4 hours; using gold nanoparticles with a diameter of 10 nm, the gold electrode modified with the capture probe DNA is immersed in the prehybridization solution at room temperature hybridization for 2 hours; the electrochemical indicator is cobalt ion (Co 2+ ), the remainder is the same as in Example 1, indicating the occurrence of the hybridization reaction by the change in the electric quantity of cobalt ions on the electrode surface before and after the hybridization. Using this method to determine the gene sequence containing 38 bases, the linear concentration range is 0.5 pmol L -1 ~10pmol·L -1 .

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Abstract

The invention describes a kind of DNA electrochemistry sensor, which comprises gold pole, complementary DNA, gold nano-particles and electrochemistry indicator. It is characterized in that one segment of complementary DNA is assembled onto the gold pole as capture probe, and the other segment as detection probe is integrated with the gold nano-particles to be the carrier of electrochemistry indicator. The preparation method is disclosed. By means of 'sandwich' hybridization method, the surface of the pole has high DNA chains with negative charge and The VIII family transition metal ions with positive charge was introduced to combine with DNA as electrochemistry indicator. The DNA hybridization reaction is indicated by detecting the electrochemistry numerical change (electric current or quantity) of the pole surface. Because of the amplification effect of the gold nano-particles, the sensor can detect trace DNA with high selectivity and the gold pole modified with DNA can be used repeatedly.

Description

technical field [0001] The invention relates to the field of bioelectrochemical sensors and preparation thereof, in particular to a deoxyribose nucleic acid (DNA) electrochemical sensor and a preparation method thereof. Background technique [0002] Genetic diagnosis is a method of diagnosing diseases by directly detecting the presence or defect of genes. The detection object of genetic diagnosis is deoxyribonucleic acid or ribonucleic acid (RNA). In terms of the methodology of genetic diagnosis, restriction endonuclease zymography analysis, nucleic acid molecular hybridization, restriction fragment length polymorphism linkage analysis, polymerase chain reaction (PCR), and DNA sensors developed in recent years have been established successively. and DNA chip technology. Labeled nucleic acid hybridization detection technology has been widely used in biology, medicine, environmental science and other related fields, but the experimental process is time-consuming and laboriou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 樊春海张炯宋世平
Owner JIANGSU WUZHONG HIGH & NEW TECH IND
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