A kind of construction method of highly stable nucleic acid sensor

A nucleic acid sensor and construction method technology, applied in the construction of high-stability nucleic acid sensors, and the construction of nucleic acid sensors, can solve problems such as easy shedding and instability, and achieve the effects of good stability, mild conditions and short construction time.

Inactive Publication Date: 2019-05-21
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The chemical reaction immobilization method is a chemical reaction between the substrate surface and the reactant. Typically, the mercapto group is combined with the gold electrode and fixed on the electrode surface. This method is unstable and easy to fall off during the electrochemical test.

Method used

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  • A kind of construction method of highly stable nucleic acid sensor
  • A kind of construction method of highly stable nucleic acid sensor
  • A kind of construction method of highly stable nucleic acid sensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] (1) Fix the Au-C bond on the electrode

[0035] First mix 10 μL of 50 μM 4-[(trimethylsilyl)ethynyl]benzoic acid THF solution with 100 μL of 20 μM tetrabutylammonium fluoride THF solution, take 55 μL into a small centrifuge tube, buckle it upside down on the electrode, Soak the electrode completely in the mixed solution. Place it at room temperature in the dark for 1h to form Au-C bonds.

[0036] (2) Formation of double-stranded DNA (dsDNA)

[0037] Prepare 50 μM DNA1Tris-HCl solution and 10 μM DNA2Tris-HCl solution respectively, mix the two solutions at 37°C, and react for 2 hours. Because the two DNAs are completely complementary, dsDNA can be obtained.

[0038] (3) Connection of dsDNA to electrode surface

[0039] The electrode was rinsed with tetrahydrofuran THF. Mix the dsDNA solution with equal volumes of 100 μM EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) and 100 μM NHS (N-hydroxysuccinimide) solution, and take in In a small centrifuge t...

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Abstract

The invention discloses a method for constructing a high-stability nucleic acid sensor. The method comprises the following steps: soaking a gold electrode in a tetrahydrofuran solution containing 4-[(trimethylsilyl) ethynyl] benzoic acid and tetrabutylammonium fluoride, and reacting in a shading room-temperature environment to obtain a 4-carboxy phenyl ethynyl modified gold electrode; carrying out a specific reaction on DNA1 and DNA2 to obtain dsDNA; carrying out an amidation reaction on the 4-carboxy phenyl ethynyl modified gold electrode and the dsDNA, and closing the surface of the gold electrode by adopting an MCH solution, thereby obtaining an adapter sensor with high stability and rapid electron transfer. Moreover, the method is simple and low in time consumption, thereby facilitating popularization and application.

Description

technical field [0001] The invention relates to a construction method of a nucleic acid sensor, in particular to a construction method of a high-stability nucleic acid sensor, and belongs to the technical field of biosensing. Background technique [0002] Electrochemical biosensors immobilize single-stranded DNA and oligonucleotide probes on the electrode surface, and use base pairing with complementary DNA strands or oligonucleotides in the sample solution for identification. Hybridization is monitored by detecting changes in the current of an electroactive indicator, or changes in electrical and interfacial properties due to the formation of a double helix structure. During the development of aptasensors, the immobilization of aptamers plays a very important role in the performance of the whole sensor. The stability, binding force and specificity to target molecules of aptasensors all depend on the immobilization technology of aptamers on the electrode, that is, the surfa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327
CPCG01N27/3275
Inventor 向娟皮晓梅曾妮
Owner CENT SOUTH UNIV
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