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DNA electrochemical biosensor and preparation method thereof

A biosensor, electrochemical technology, applied in the field of material preparation, detection analysis, nucleic acid detection, can solve problems such as unsatisfactory selectivity, and achieve the effect of reducing false positive reactions, high specificity, and high binding

Inactive Publication Date: 2015-04-29
UNIV OF SCI & TECH BEIJING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Electroactive intercalators are a well-studied class of hybridization indicators, however, they tend to bind both single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA), so their selectivity is not ideal

Method used

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  • DNA electrochemical biosensor and preparation method thereof
  • DNA electrochemical biosensor and preparation method thereof
  • DNA electrochemical biosensor and preparation method thereof

Examples

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Embodiment 1

[0048] 1. Pretreatment of the gold electrode: Polish the gold electrode with 0.3 μm and 50 nm polishing powder for 5 minutes, and then perform ultrasonic cleaning with acetone, ethanol, and ultrapure water respectively, and the ultrasonic cleaning time is 3 minutes. Then put the above electrode into 0.1mol / L dilute H 2 SO 4 In the process, it is thoroughly cleaned by electrochemical methods to obtain clean gold electrodes.

[0049]2. Preparation of the modified gold electrode: Soak the pretreated gold electrode in 50nmol / L mercapto-ssDNA solution (prepared with pH 7.0 phosphate buffer solution) for 16h, then soak the electrode in 1mmol / L mercapto In ethanol solution for 1 h, the ssDNA probe modified gold electrode was obtained.

[0050] 3. Preparation of DNA electrochemical sensor: Soak the modified gold electrode in 0.5mg / mL conductive nanomaterial solution for 0.5h to react.

[0051] 4. Soak the electrode obtained in the above 3 in the target strand DNA T 1 The solution ...

Embodiment example 2

[0059] 1. Pretreatment of the gold electrode: Polish the gold electrode with 0.3 μm and 50 nm polishing powder for 5 minutes, and then perform ultrasonic cleaning with acetone, ethanol, and ultrapure water respectively, and the ultrasonic cleaning time is 5 minutes respectively. Then put the above electrode into 0.3mol / L dilute H 2 SO 4 In the process, the surface is thoroughly cleaned by electrochemical methods to obtain clean gold electrodes.

[0060] 2. Preparation of ssDNA probe modified gold electrode: Soak the pretreated gold electrode in 50nmol / L thiolated ssDNA solution (prepared with pH 7.0 phosphate buffer solution) for 24h, and then soak the electrode in 1mmol / L L of mercaptohexanol solution for 1.5h to obtain a modified gold electrode.

[0061] 3. Preparation of DNA electrochemical sensor: Soak the modified gold electrode in 0.5mg / mL conductive nanomaterial solution for 0.5h to react.

[0062] 4. Soak the electrode obtained in the above 3 in the complementary st...

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Abstract

The invention discloses a DNA electrochemical biosensor and a preparation method thereof. The DNA electrochemical biosensor comprises a gold electrode, a complementary DNA complementary with a target DNA sequence, conductive nanoparticles, and a soluble electrochemical active reagent, wherein a complementary single chain DNA, as a capturing probe, is assembled on the gold electrode; the single chain DNA and the conductive nanoparticles are combined as an electrochemical signal conversion unit for realizing electrochemical response to a DNA hybridizing event. By the preparation method, the sulfhydrylated DNA modified gold electrode adsorbed with the conductive nanoparticles is used as a signal conversion electrode, and through electrode reaction of the soluble electrochemical active reagent, based on base complementing and pairing between the DNA sequences, electrochemical response and detection of the target DNA is realized. The DNA electrochemical biosensor is simple in structure, simple and convenient in preparation process and high in sensibility, can be applied to quick detection of DNA molecules, and has the characteristic of reusability.

Description

technical field [0001] The invention belongs to the technical field of material preparation and detection and analysis, and relates to a sensor and a manufacturing method thereof, in particular to a DNA electrochemical biosensor and a manufacturing method thereof, which are applied in the technical field of nucleic acid detection. Background technique [0002] Deoxyribonucleic acid (DNA) is the bearer of genetic information, and the variation of base sequences in DNA molecules is related to many genetic diseases of human beings. Therefore, the analysis of specific sequence of DNA and the detection of base mutations in DNA chains have far-reaching significance in gene screening, early diagnosis and treatment of genetic diseases. DNA biosensor is an important method for structural analysis and detection of nucleic acid. Among the many hybridization detection methods, electrochemical analysis technology has the characteristics of simple instrument, low price, fast and accurate ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
Inventor 王媛媛苏磊佟莹杨涵焜舒桐龚伟张学记
Owner UNIV OF SCI & TECH BEIJING
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