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Photo-electro-chemical method for detecting nucleic acid

A photoelectrochemical and nucleic acid technology, applied in biochemical equipment and methods, measuring devices, scientific instruments, etc., can solve the problems of limited selection range and limited detection range of signal molecules

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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the method can directly detect the oxidation-reduction reaction of nucleic acid bases without labeling, the detection range of this method is limited, and the method uses voltage on the electrode to excite signal molecules, so that the selection range of signal molecules is limited

Method used

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  • Photo-electro-chemical method for detecting nucleic acid

Examples

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Effect test

preparation example Construction

[0064] 1. Preparation of Nano-SnO Semiconductor Electrode

[0065] The indium tin oxide conductive glass was ultrasonically washed with detergent (15 minutes), pure water (2 minutes, twice), acetone (5 minutes), isopropanol (5 minutes) and pure water (10 minutes, twice) , blow dry, take the tin dioxide colloidal solution with a particle size of 15nm and evenly spread it on the ITO electrode plate. After drying, sintering at 450° C. for 1 hour, cutting into small electrode pieces of 0.5 cm×2.5 cm to obtain nanometer tin dioxide semiconductor electrodes.

[0066] 2. Preparation of ruthenium complex-labeled avidin

[0067] Put 2mg of avidin (concentration: 10mg / mL, volume: 200uL, solvent: 30mM phosphate buffer, pH 8.0) into 30uL containing 0.7mg ruthenium bis(2,2'-bipyridine) (4'-methyl -4-carboxy-2,2'-bipyridine)-succinamide ester in anhydrous N,N-dimethylamide solution, and shake the reaction for 90 minutes in the dark. The unlabeled ruthenium complex is separated by centrif...

Embodiment 1

[0071] Photocurrent measurement

[0072] Photocurrent was measured on a CHI 630 Electrochemical Analyzer in time-based mode. The light source is a 20mW laser with a wavelength of 470nm. The micro photoelectrochemical cell is made of polished glass, the reference electrode is Ag / AgCl, and the counter electrode is platinum electrode. The light beam is injected vertically into the cell, where it hits a tin dioxide electrode on its back side. Manually move the opaque cardboard to make the light beam irradiate the electrodes regularly and intermittently. The ruthenium bipyridine complex has an absorption maximum at 470nm.

[0073] Such as figure 1 As shown, the nano-tin oxide electrode adsorbed with Ru-Avidin was placed in a photoelectrochemical cell in 20 mM phosphate buffer, pH 5.5. Block the light beam with a cardboard at the beginning of the measurement, and pull the cardboard away every 10 seconds. It can be seen that the photoelectrochemical substance ruthenium complex ...

Embodiment 2

[0076] Detection of guanine-catalyzed photocurrents

[0077] Dilute guanine nucleotides with 20mM phosphate buffer pH 5.5 to the following concentration gradients: 1μg / mL, 5μg / mL, 10μg / mL, 20μg / mL and 50μg / mL. Add the diluted guanylic acid solution into the photoelectrochemical cell for photoelectric detection, and obtain photocurrents catalyzed by different concentrations of guanylic acid. The result is as image 3 Shown (a is 0 μg / mL; b is 1 μg / mL; c is 5 μg / mL; d is 10 μg / mL; e is 20 μg / mL; f is 50 μg / mL). The results of this experiment show that the base guanine has a strong catalytic effect on the photocurrent generated by the photoelectrochemical substance, and the higher the concentration, the stronger the catalytic effect.

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Abstract

The invention relates to a method for detecting nucleic acid alkali group with optical electric chemical technique, wherein said method comprises: a, contacting the tested sample and the material with optical electric chemical activity; b, evaluating the oxidization and reduction reaction between the nucleic acid alkali group and said optical electric chemical material, that using light to transform the optical electric chemical material into exciting state with existed electrode, and processing oxidization reduction reaction with nucleic acid alkali group when the excited material loses electrons, to evaluate the current generated in oxidization reduction reaction; c, based on said current, analyzing the sample qualitative or quantitatively. The invention can be used in biological detection, as nucleic acid cross detection, nucleic hurt detection and chemical material gene toxicity detection.

Description

technical field [0001] The invention relates to the technical field of nucleic acid detection, in particular to a method for qualitatively and quantitatively detecting nucleic acid hybridization and damage using photoelectrochemistry. technical background [0002] It is known that human diseases are directly or indirectly related to genes. Therefore, there is an increasingly urgent demand for highly sensitive and highly selective rapid detection methods for nucleic acids, which are of great importance for gene research, drug screening, clinical diagnosis and Ecotoxicology research, toxicity screening of new compounds, and monitoring of biological pollutants in the environment all have practical significance. [0003] Many detection methods have been used to detect nucleic acids, for example, Southern blotting, Northern blotting and PCR techniques commonly used in nucleic acid detection at present. A common point in the operation of these methods is that unhybridized probes a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50G01N27/00C12Q1/68
Inventor 郭良宏梁敏敏
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