Colorimetric detection method based on nanometer-gold and nucleic acid structure and kit thereof

A detection method, nano-gold technology, which is applied in the direction of material analysis by observing the influence of chemical indicators, and analysis by making materials undergo chemical reactions, can solve problems such as inapplicability, and achieve low cost, high specificity, The effect of improving sensitivity

Inactive Publication Date: 2010-09-15
苏州市长三角系统生物交叉科学研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method only detects specific DNA, and is not suitable for the detection of any target substance

Method used

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  • Colorimetric detection method based on nanometer-gold and nucleic acid structure and kit thereof
  • Colorimetric detection method based on nanometer-gold and nucleic acid structure and kit thereof
  • Colorimetric detection method based on nanometer-gold and nucleic acid structure and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The detection of embodiment 1 cocaine

[0041] Steps: Take 2 μL of cocaine-nucleic acid aptamer solution with a concentration of 100 μM and 2 μL of an aqueous solution of cocaine hydrochloride with a concentration of 0.1 to 10 mM, add 18 μL of buffer solution (25 mM Tris, pH 8.2, 0.6 M NaCl) and mix ( The final concentration of DNA is 10 μM, the final concentration of cocaine is 10-1000 μM), and the reaction is carried out at room temperature for 30 min. 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 reaction solution and add 100 μL of the nano-gold solution (13nm, 3.5nM) prepared above. ) (final concentration about 50mM). Observe the color change of gold nanoparticles in the experimental group and the control group and record the ultraviolet-visible spectrum.

[0042] Result...

Embodiment 2

[0043] The detection of embodiment 2ATP

[0044] Steps: Take 2 μL of ATP-nucleic acid aptamer solution with a concentration of 10 μM and 2 μL of an aqueous solution of ATP with a concentration of 0.01 to 1 mM, add 18 μL of buffer solution (10 mM Tris, pH 8.2, 0.3 M NaCl) and mix (final DNA concentration 1 μM, the final concentration of ATP is 1-100 μM), and reacted at room temperature for 15 minutes. 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 10 μL of the nano-gold solution (20nm, 2nM) prepared above, and react at room temperature for 5 minutes, then add 10 μL of 0.2M PBS (10mM PB, pH7.0, 0.2M NaCl) (PBS The final concentration is approximately 100 mM). The color changes and ultraviolet-visible spectra of the gold nanoparticles in the experimental group and the control group were recorded r...

Embodiment 3

[0046] Embodiment 3 is to the detection of divalent mercury ion

[0047] Step: Take 2 μL of Hg with a concentration of 100 μM 2+ - MSO solution, add 2 μL of Hg with a concentration of 0.1-2 mM 2+ aqueous solution (the reaction concentration of MSO is 50 μM, the reaction concentration of the target substance is 50-1000 μM, the reaction buffer solution and the ionic strength are both 0), react at room temperature for 1 min. 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 Ca2+ , 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 2 μL of the above reaction solution and add 100 μL of the nano-gold solution (10nm, 100nM) prepared above, and react at 37°C for 1min, then add 60μL of 1M NaNO 3 (Final concentration of salt is approximately 600 mM). Observe the color change o...

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Abstract

The invention discloses a colorimetric detection method based on a nanometer-gold and nucleic acid structure and a kit thereof. The colorimetric detection method comprises the following steps of: (1) mixing specific DNA which can be specifically combined with the target molecules with a solution to be detected for reaction; (2) mixing the reaction solution which is obtained in the step (1) with a nanometer-gold solution of which the number of moles is 0.01 to 1 time that of the specific DNA for reaction; and (3) adding a high-salt solution with the reaction concentration between 10 and 600 mM, and observing color change of the solution. In the operation process of the invention, DNA is not needed to be marked, expensive equipment is not needed need, and the existence of the target molecules can be determined only by observing the color change of the nanometer-gold solution. Therefore, the invention is simple, convenient and fast, has low cost, high specificity and high sensitive detection of any target molecules, is especially suitable for open-air analysis and high-throughput screening, and has wide application range.

Description

technical field [0001] The invention particularly relates to a colorimetric detection method based on nano gold and nucleic acid structure and a kit thereof. 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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78
Inventor 樊春海王丽华
Owner 苏州市长三角系统生物交叉科学研究院有限公司
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