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Target molecule detecting method based on nanometer aurum and nucleic acid structure

A detection method and technology of target molecules, which are applied in the determination/inspection of microorganisms, biochemical equipment and methods, and analysis through chemical reactions of materials, etc., to achieve the effects of wide application range, rapid detection and high specificity

Inactive Publication Date: 2009-05-06
SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The technical problem to be solved by the present invention is to overcome the defect that different target molecules need to be designed differently in the existing method for detecting target DNA by nano gold, and to provide a fast, high specificity, high sensitivity, general-purpose The method of nano-gold detection target substance can easily realize the detection of any target substance

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  • Target molecule detecting method based on nanometer aurum and nucleic acid structure
  • Target molecule detecting method based on nanometer aurum and nucleic acid structure

Examples

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Effect test

Embodiment 1

[0036] Example 1 Detection of cocaine

[0037] Steps: Take 2 μL cocaine aptamer solution with a concentration of 100 μM and 2 μL cDNA solution with a concentration of 100 μM, add 14 μL buffer solution (25 mM Tris, pH 8.2, 0.3 M NaCl), heat to 85 ° C and slowly cool to Allow it to fully hybridize to form double-stranded probes at room temperature. Then, 2 μL of cocaine hydrochloride aqueous solution with a concentration of 0.01-1 mM was added to the hybridization solution, and the mixture was reacted at room temperature for 30 minutes to fully react. At the same time, the group without adding cocaine and adding 2 μL of water was used as a blank, and the two groups of adding 2 μL of 1 mM benzoylecgonine (N1) and ecgonine methyl ester (N2) were used as controls. Then take 2 μL of the above-mentioned reaction solution and add 100 μL of nano-gold solution (13nm, 3.5nM), react for 5min, use the above-mentioned buffer solution (25mM Tris, pH8.2, 0.3M NaCl) to make up the total volum...

Embodiment 2

[0039] The detection of embodiment 2 ATP

[0040] Steps: Take 2 μL of ATP aptamer solution with a concentration of 100 μM and 2 μL of cDNA solution with a concentration of 100 μM, add 14 μL of buffer solution (25 mM Tris, pH 8.2, 0.15 M NaCl), heat to 85 ° C and slowly cool to Allow it to fully hybridize to form double-stranded probes at room temperature. Then, 2 μL of an aqueous solution of ATP with a concentration of 0.01-1 mM was added to the hybridization solution, and the mixture was reacted at room temperature for 30 minutes to fully react. At the same time, a group without adding ATP and adding 2 μL of water was used as a blank, and a group of adding 2 μL of 1 mM CTP, UTP, and GTP was used as a control. Then take 2 μL of the above-mentioned reaction solution and add 100 μL of nano-gold solution (20nm, 1nM), react for 5 minutes, use the above-mentioned buffer solution (25mM Tris, pH8.2, 0.15M NaCl) to make up the total volume of 0.2mL, record the experimental group and...

Embodiment 3

[0042]Example 3 Detection of divalent mercury ions (1)

[0043] Steps: Take 1 μL of MSO solution with a concentration of 1 μM and 1 μL of cDNA solution with a concentration of 1 μM, add 16 μL of buffer solution (10 mM arsine-sodium arsine, pH 6.8, 0.3 M NaCl), and react for 10 minutes at room temperature. It hybridizes sufficiently to form a double-stranded probe. Then add 2 μL of Hg with a concentration of 100 nM to 0.1 mM to the hybridization solution. 2+ aqueous solution, and reacted at room temperature for 5 minutes to make it fully reacted. without adding Hg 2+ , add 2 μL of water to a group as a blank experiment. In addition, selectivity analysis was carried out through two groups of experiments, one group added 2 μL of 25 mM Ca 2+ , Mg 2+ The other group added 0.5mM mixed ions (Fe 2+ , Cu 2+ ,Co 2+ , Mn 2+ , Ni 2+ , Zn 2+ , Cd 2+ ). Then take 5 μL of the above-mentioned reaction solution and add 5 μL of nano-gold solution (10nm, 100nM) respectively, after r...

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Abstract

The invention discloses a detection method of target molecules, which is based on manometer gold and a nucleic acid structure and comprises the sequential steps: (1), hybridizing specific DNA (Deoxyribonucleic Acid) with cDNA (Complementary Deoxyribonucleic Acid) in fluorescence labeling to form a double-stranded capture probe, wherein the specific DNA can be in specificity combination with the target molecules; (2), adding a solution to be detected for reaction; (3), adding a manometer gold solution for reaction and recording a fluorescence spectrum of the reaction solution. The detection method of the invention has commonality and wide range of application so that detection objects can be any target molecule. The invention can be utilized to conveniently realize the rapid, sensitive and selective detection of the target molecules.

Description

technical field [0001] The invention particularly relates to a target molecule detection method based on nano gold and nucleic acid structure. Background technique [0002] Nucleic acid aptamer (aptamer) refers to the short single-stranded oligonucleotide pairing obtained by screening from a random oligonucleotide library using the SELEX (systematic evolution of ligands by exponential enrichment) in vitro screening technology established at the end of the last century. group, it can specifically bind to the target molecule, thereby undergoing a conformational change itself. [0003] Through in vitro screening technology, nucleic acid aptamers of any substance can theoretically be screened, coupled with the technical characteristics of high-throughput screening and the characteristics of precise recognition of nucleic acid aptamers, easy in vitro synthesis and modification, etc., making nucleic acid aptamers in analytical chemistry and It has broad application prospects in b...

Claims

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Application Information

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IPC IPC(8): G01N21/76C12Q1/68
Inventor 樊春海王丽华张娟
Owner SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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