Method and device for fixing and assembling DNA and markers

An assembly method and a technology of markers, which are applied in the fields of analysis and detection and material science, can solve the problems of cumbersome operation, large background signal influence, and unfavorable preparation of DNA probes, etc., and achieve the effects of less reagent consumption, simple method, and low cost

Inactive Publication Date: 2008-07-23
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is easy to operate, but the presence of a large amount of protein affects the detection sensitivity and selectivity; the LB membrane method can obtain ordered monomolecular membranes, but the assembled membrane is unstable, which is not conducive to the preparation of higher-density DNA probes; The chemical polymerization fixation method usually does not need to be labeled again, and the operation is simple, but the background signal has a great influence
The covalent bonding method is also widely used for the immobilization of DNA probes on the carrier, and there are often disadvantages such as cumbersome operation.

Method used

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  • Method and device for fixing and assembling DNA and markers
  • Method and device for fixing and assembling DNA and markers
  • Method and device for fixing and assembling DNA and markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The assembly of embodiment 1 single-stranded DNA and marker on ITO conductive glass

[0036] The selected single-stranded DNA sequence structure is AACCAACACA, and the concentration is 6 μmol L -1 . The label is ruthenium complex [Ru(bpy) 2 tatp]Cl 2 , the concentration is 0.2mmolL -1 . Prepare the mixed solution of the two, take 0.4mL and add it to a chamber of the multi-channel electrolytic cell, and perform continuous differential pulse voltammetry scanning in the range of 0.2V to 1.5V. After scanning 21 times, record the differential pulse voltammetry spectrum as shown in Figure 4. It can be seen from the figure that as the number of differential pulse voltammetry scans increases, the assembly peak current gradually increases, and the assembly amount of single-stranded DNA sequences and markers on the ITO surface can be controlled by controlling the number of differential pulse voltammetry scans.

Embodiment 2

[0037] Example 2: Assembly of double-stranded DNA and markers on ITO conductive glass

[0038] The selected double-stranded DNA is calf thymus DNA, the concentration is 0.2mmolL -1 . The label is ruthenium complex [Ru(bpy) 2 tatp]Cl 2 , the concentration is 0.2mmolL -1 . Prepare the mixed solution of the two, take 0.4mL and add it to a chamber of the multi-channel electrolytic cell, and perform continuous differential pulse voltammetry scanning in the range of 0.2V to 1.5V. After scanning 21 times, record its differential pulse voltammetry spectrum, see the attached picture 5. It can be seen from the figure that as the number of differential pulse voltammetry scans increases, the assembly peak current gradually increases, and the assembly amount of single-stranded DNA sequences and markers on the ITO surface can be controlled by controlling the number of differential pulse voltammetry scans.

Embodiment 3

[0039] Example 3: Multi-channel assembly of DNA and markers on ITO conductive glass

[0040] The selected double-stranded DNA is calf thymus DNA, and the concentrations are 0, 6, 12, 18, 24, 30 μmol L -1 . The label is ruthenium complex [Ru(bpy) 2 tatp]Cl 2 , the concentration was fixed at 0.2mmol L -1 . Prepare six kinds of mixed solutions of the two respectively, take 0.4mL each and add them to the six chambers of the multi-channel electrolytic cell, conduct continuous differential pulse voltammetry scanning in the range of 0.2V to 1.3V, and record the differential pulse voltage after scanning 21 times For the voltammogram, see Figure 6. It can be seen from the figure that the assembly peak current increases with the increase of DNA concentration, and this method can be used to quantitatively measure the concentration of DNA.

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Abstract

The invention discloses a method for immobilizing and assembling a DNA and a marker, which is characterized in that a testing solution comprising the DNA and the marker is prepared and filled in an electrolytic tank to do a continuous differentiating impulse volt-ampere scanning, completing the electrochemical assembly of the DNA and the marker on a conductive glass surface. The invention simultaneously discloses a multi-channel electrolytic tank to complete the process. The method for immobilizing and assembling a DNA and the marker has the advantages of convenient operation, less reagent consumption, low cost, real-time monitoring for the assembly amount of the DNA and the marker on the ITO surface, and synchronous implementation of assembly.

Description

technical field [0001] The invention relates to the technical fields of analysis and detection and material science, and mainly relates to an electrochemical array assembly technology of DNA and markers. Background technique [0002] As the genetic material of organisms, DNA has a unique π system. Using the electrostatic interaction, groove surface binding or intercalation between polypyridine ruthenium complexes and DNA can construct molecular assemblies with specific uses, thus providing new ideas for the development of gene chips, gene sensors, DNA computers and other fields. [0003] At present, the methods of immobilizing DNA mainly include chemical adsorption method, self-assembled membrane method, synthesis method, biotin-avidin reaction method, Langmuir-Blodgett (LB) membrane method, polymer membrane method and covalent bonding method. The immobilization method based on chemical adsorption is simple to operate, but it is easy to desorb, and due to multi-point adsorp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12M1/36C12M1/42
Inventor 李红姚夙
Owner SOUTH CHINA NORMAL UNIVERSITY
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