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Fluorescence detection method for copper ion based on click chemistry

A technology of fluorescence detection and click chemistry, which is applied in the field of chemical analysis and detection, can solve the problems of low sensitivity and cumbersome detection steps, and achieve the effects of strong anti-interference ability, reducing the possibility of false positive signals, and mild reaction conditions

Active Publication Date: 2014-08-27
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to solve the problem that the existing methods for detecting copper ions based on click chemistry fluorescence need to add substances such as heavy metals or toxic dyes, and the detection steps are cumbersome and low in sensitivity, and provide a method for fluorescence detection of copper ions based on click chemistry

Method used

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  • Fluorescence detection method for copper ion based on click chemistry
  • Fluorescence detection method for copper ion based on click chemistry
  • Fluorescence detection method for copper ion based on click chemistry

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Embodiment 1

[0042] Step 1. The short single-stranded nucleic acid modified with an azide group, as shown in SEQ ID No: 1 (5'-CTA AAT TCC AA-azide-3'), was modified with a short alkynyl group. Single-stranded nucleic acid, as shown in SEQ ID No: 5 (5'-alkynyl-GAA ACT GAT AG-3'), and nucleic acid as ligation template, such as SEQ ID No: 9 (5'-FAM-TCG CTA TCA GTT TCT TGG AAT TTA GCG A-Dabcyl-3') were respectively dissolved in 20 mM Tris-HCl (pH 7.4) buffer solution to prepare nucleic acid solutions with a concentration of 6 μM;

[0043] Step 2. Mix the short single-stranded nucleic acid solution modified with azide group, the short single-stranded nucleic acid solution modified with alkynyl group, and the nucleic acid solution as a connection template in a ratio of 3:3:1 to form a mixed solution. solution, and then add 500 μM ascorbic acid, water and 20 mM Tris-HCl (pH 7.4) buffer solution to the mixed solution to form a detection system, in which the short single-stranded nucleic acid solut...

Embodiment 2

[0048] Step 1. The short single-stranded nucleic acid modified with an azide group, as shown in SEQ ID No: 1 (5'-CTA AAT TCC AA-azide-3'), was modified with a short alkynyl group. Single-stranded nucleic acid, as shown in SEQ ID No: 5 (5'-alkynyl-GAA ACT GAT AG-3'), and nucleic acid as ligation template, such as SEQ ID No: 9 (5'-FAM-TCG CTA TCA GTT TCT TGG AAT TTA GCG A-Dabcyl-3') were respectively dissolved in 20 mM Tris-HCl (pH 7.4) buffer solution to prepare nucleic acid solutions with a concentration of 6 μM;

[0049] Step 2. Mix the short single-stranded nucleic acid solution modified with azide group, the short single-stranded nucleic acid solution modified with alkynyl group, and the nucleic acid solution as a connection template in a ratio of 3:3:1 to form a mixed solution. solution, and then add 500 μM ascorbic acid, water and 20 mM Tris-HCl (pH 7.4) buffer solution to the mixed solution to form a detection system, in which the short single-stranded nucleic acid solut...

Embodiment 3

[0055] Step 1. The short single-stranded nucleic acid modified with an azide group, as shown in SEQ ID No: 1 (5'-CTA AAT TCC AA-azide-3'), was modified with a short alkynyl group. Single-stranded nucleic acid, as shown in SEQ ID No: 5 (5'-alkynyl-GAA ACT GAT AG-3'), and nucleic acid as ligation template, such as SEQ ID No: 9 (5'-FAM-TCG CTA TCA GTT TCT TGG AAT TTA GCG A-Dabcyl-3') were respectively dissolved in 20 mM Tris-HCl (pH 7.4) buffer solution to prepare nucleic acid solutions with a concentration of 6 μM;

[0056] Step 2. Mix the short single-stranded nucleic acid solution modified with azide group, the short single-stranded nucleic acid solution modified with alkynyl group, and the nucleic acid solution as a connection template in a ratio of 3:3:1 to form a mixed solution. solution, and then add 500 μM ascorbic acid, water and 20 mM Tris-HCl (pH 7.4) buffer solution to the mixed solution to form a detection system, in which the short single-stranded nucleic acid solut...

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Abstract

The invention provides a fluorescence detection method for copper ions based on click chemistry and belongs to the technical field of chemical analysis and detection. A short single chain nucleic acid which is modified by an azide group, a short single chain nucleic acid which is modified by an alkynyl group and a nucleic acid which is used as a linking template are respectively dissolved in a Tris-HCl buffer solution to prepare a nucleic acid solution and form a mixed solution. Ascorbic acid, water and a Tris-HCl buffer solution are added in the mixed solution to form a detection system. A to-be-detected sample is added to the detection system. A fluorescence detection process is carried out to the fluorescence intensity of the detection system. Copper ion concentration of the corresponding to-be-detected sample is calculated with a known copper ion concentration being an abscissa and a corresponding fluorescence intensity value being an ordinate. The fluorescence detection method employs a linking reaction of nucleic acids based on the click chemistry, is simple and quick, is good in stability and high in sensitivity, and has a good linear response in a range from 0 to 40 nM and a detection limit as low as 0.023 nM.

Description

technical field [0001] The invention belongs to the field of chemical analysis and detection, and in particular relates to a click chemistry-based copper ion fluorescence detection method. Background technique [0002] Copper is one of the indispensable trace elements to maintain human health and normal functions of other organisms. Copper plays a very important role in physiological processes by acting as a cofactor and / or structural component of enzymes and other proteins, and is closely related to the development of the blood, reproductive system, and bones. The lack of copper in the human body will cause the growth retardation of the human body and various diseases related to copper deficiency, such as celiac disease, anemia, cardiovascular injury, spinal myelination and so on. Therefore, the World Health Organization (WHO) and the Food and Agriculture Organization of the United Nations (FAO) recommend that the daily intake of copper should not be lower than 0.5mg / kg. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 于聪王方远李永新张青峰陈健周会鹏
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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