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Electrochemical sensor based on DNA configuration transformation

A configuration conversion and electrochemical technology, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., to achieve the effect of reducing complexity

Inactive Publication Date: 2016-07-20
NANJING UNIV OF POSTS & TELECOMM
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Existing electrochemical DNA sensors are limited to one configuration change and detect one target to be tested. However, in practical applications, we often need to detect two or more targets at the same time, so the application of traditional sensors has encountered great difficulties. limitations

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  • Electrochemical sensor based on DNA configuration transformation
  • Electrochemical sensor based on DNA configuration transformation
  • Electrochemical sensor based on DNA configuration transformation

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Embodiment Construction

[0026] The capture probe DNA is 5'-thiol-TTTACCTGGACCTTCCTCCGCAATACTCCCCCAGGTTTT-MB-3', the aptamer DNA is 5'-ACCTGGGGGAGTATTGCGGAGGAAGGT-3', and the aptamer and neck loop DNA are selected according to the target.

[0027] Construction of Neckloop DNA Electrochemical Sensor:

[0028] Soak the electrode in sodium borohydride solution for 1 hour, after rinsing, polish the electrode with 0.3μm and 0.05μm alumina powder in turn, use ethanol and ultrapure water for 2 minutes, in 0.5mol / L NaOH solution, from -0.35 to 1.5V Cycle scan 20, use 0.5mol / LH in electrochemical workstation Cell 2 SO 4 Solution processing, finally in new 0.5mol / LH 2 SO 4 The solution scans out the characteristic peaks of the gold electrode.

[0029] Disperse the capture probe of the hairpin structure in a Tris-TE buffer solution with a pH of 7.0 (preparation method: prepare 20mmol / L Tris-base, add 140mmol / LNaCl and 5mmol / LMgCl 2, , adjusted the pH to 7.0 with 0.2 mol / L HCl to obtain a diluted Tris-HCL bu...

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Abstract

The invention discloses an electrochemical sensor based on DNA configuration transformation.The electrochemical sensor comprises an electrode and a capturing probe DNA.The capturing probe DNA is of a nuchal-ring single-chain structure, the 3' end of a DNA chain is used for modifying methylene blue, the 5' end of the DNA chain is used for modifying sulfydryl, and the capturing probe DNA stands on the surface of the electrode under the interaction between the sulfydryl and the electrode.According to the electrochemical sensor, the electrochemical indicator methylene blue (MB) is modified to the nuchal-ring DNA probe; after target DNA and ATP are reacted with the probe DNA, the configuration of the probe DNA is changed, an electrochemical signal is changed, and the target DNA and the ATP are detected at the same time without replacing the sensor.In other words, two components are detected with one nuchal-ring DNA, and the complexity of an experiment is lowered.On the basis, it can be further expanded that selective or simultaneous detection or any two or more target molecules is achieved with one DNA probe.

Description

technical field [0001] The invention belongs to the field of preparation and application of electrochemical biosensors, and relates to a preparation method and application of an electrochemical sensor based on neck loop DNA. Background technique [0002] The electrochemical research of DNA began in the 1960s, and the early work mainly focused on the basic electrochemical behavior of DNA. With the development of polarography, other electroanalytical chemical methods are also used in the study of DNA, and its scope has been expanded to the study of DNA structure and morphology, DNA probes and sensors. DNA biosensors are playing an increasingly important role in health and epidemic prevention and disease diagnosis because of their rapidity, sensitivity, simple operation and equipment, especially their molecular recognition function, and are used in gene diagnosis, treatment, and infectious disease detection. Made a lot of contributions. [0003] In terms of DNA hybridization ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/48
CPCG01N27/3277G01N27/48
Inventor 樊春海苏邵刘巍汪联辉晁洁邹敏张池曹文芳韩小彦卢在伟
Owner NANJING UNIV OF POSTS & TELECOMM
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