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dsDNA (double-stranded deoxyribonucleic acid) high-sensitivity detection method based on monochrome fluorescence off-on switching system

A sensitive detection and switching system technology, applied in the fields of chemistry and biomedicine, can solve the problems of low detection sensitivity and achieve high sensitivity, good specificity and low cost

Inactive Publication Date: 2014-01-22
BEIJING INSTITUTE OF TECHNOLOGYGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the problem of low detection sensitivity of the existing technology for detecting dsDNA, and to provide a high-sensitivity detection method for dsDNA based on a monochromatic fluorescent "off-on" switch system

Method used

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  • dsDNA (double-stranded deoxyribonucleic acid) high-sensitivity detection method based on monochrome fluorescence off-on switching system
  • dsDNA (double-stranded deoxyribonucleic acid) high-sensitivity detection method based on monochrome fluorescence off-on switching system
  • dsDNA (double-stranded deoxyribonucleic acid) high-sensitivity detection method based on monochrome fluorescence off-on switching system

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Embodiment 1

[0021] A high-sensitivity detection method for dsDNA based on a monochromatic fluorescent "off-on" switch system of the present invention, such as figure 1 As shown, the specific steps are as follows:

[0022] Step 1, synthesizing CdTe quantum dots (QDs) coated with water-soluble glutathione (GSH), controlling the fluorescence emission wavelength of the quantum dots at 605nm;

[0023] Synthesis steps: 63.8mg of Te powder and 100mg of NaBH 4 Mix, then add 5mL N 2 Saturated deionized water, mixed solution in N 2 React under protection for 2h until the solution turns pale pink, and this solution is called Te precursor here.

[0024] 0.2312g (1mmol) of CdCl 2 2.5H 2 O, 0.3685g (1.2mmol) of glutathione (GSH), and 100mL of deionized water were mixed, and under magnetic stirring, NaOH was added dropwise to adjust the pH to 8-9 to obtain a mixed solution. Pass 30min N 2 After that, the precursor solution of Te was quickly injected into the mixed solution, and the N 2 Under pro...

Embodiment 2

[0035] A high-sensitivity detection method for dsDNA based on a monochromatic fluorescent "off-on" switch system of the present invention, such as figure 1 As shown, the specific steps are as follows:

[0036] Step 1, synthesizing CdTe quantum dots (QDs) coated with water-soluble glutathione (GSH), controlling the fluorescence emission wavelength of the quantum dots at 605nm;

[0037] Synthesis steps: 63.8mg of Te powder and 100mg of NaBH 4 Mix, then add 5mL N 2 Saturated deionized water, mixed solution in N 2 React under protection for 2h until the solution turns pale pink, and this solution is called Te precursor here.

[0038] 0.2312g (1mmol) of CdCl 2 2.5H 2 O, 0.3685g (1.2mmol) of glutathione (GSH), and 100mL of deionized water were mixed, and under magnetic stirring, NaOH was added dropwise to adjust the pH to 8-9 to obtain a mixed solution. Pass 30min N 2 After that, the precursor solution of Te was quickly injected into the mixed solution, and the N 2 Under pro...

Embodiment 3

[0046] A high-sensitivity detection method for dsDNA based on a monochromatic fluorescent "off-on" switch system of the present invention, such as figure 1 As shown, the specific steps are as follows:

[0047] Step 1, synthesizing CdTe quantum dots (QDs) coated with water-soluble glutathione (GSH), controlling the fluorescence emission wavelength of the quantum dots at 605nm;

[0048] Synthesis steps: 63.8mg of Te powder and 100mg of NaBH 4 Mix, then add 5mL N 2 Saturated deionized water, mixed solution in N 2 React under protection for 2h until the solution turns pale pink, and this solution is called Te precursor here.

[0049] 0.2312g (1mmol) of CdCl 2 2.5H 2 O, 0.3685g (1.2mmol) of glutathione (GSH), and 100mL of deionized water were mixed, and under magnetic stirring, NaOH was added dropwise to adjust the pH to 8-9 to obtain a mixed solution. Pass 30min N 2 After that, the precursor solution of Te was quickly injected into the mixed solution, and the N 2 Under pr...

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Abstract

The invention relates to a dsDNA (double-stranded Deoxyribonucleic Acid) high-sensitivity detection method based on a monochrome fluorescence off-on switching system and belongs to the field of chemistry and biomedicine. The dsDNA high-sensitivity detection method comprises the following steps: synthesizing CdTe quantum dots (QDs) coated by water-soluble glutathione (GSH); controlling the fluorescence-emission wavelength of the quantum dots to be 605mm; mixing the CdTe quantum dots coated by the water-soluble glutathione with a metal ruthenium coordination compound [Ru(phen)2(dppz)]2+(Ru) to enable the fluorescence of the CdTe quantum dots coated by the water-soluble glutathione to be completely quenched, so as to obtain a QDs-Ru assembling group; adding dsDNA into the QDs-Ru assembling group and gradually enhancing the monochrome fluorescence strength at the wavelength of 605mm along with gradual increase of the concentration of the dsDNA so as to rapidly and flexibly defect the dsDNA. The detection system can be used for enabling the detection limit of the dsDNA to reach 10pg / mL. Compared with the prior art in a background technology, the sensitivity is improved by 500 times; the operation is simple and rapid; the specificity is good and the cost is low; the whole process can be finished within 30 minutes.

Description

technical field [0001] The invention relates to a dsDNA high-sensitivity detection method based on a monochromatic fluorescent "off-on" switch system, which belongs to the fields of chemistry and biomedicine. Background technique [0002] As an excellent fluorescent nanomaterial, quantum dots (QDs) have many unique characteristics such as good chemical stability, high fluorescence efficiency, wide excitation spectrum, and narrow emission spectrum. They can be combined with biomolecules to prepare bioluminescent probes for biological Molecular detection and sensing. As the carrier of genetic information, DNA's rapid and highly sensitive detection is becoming more and more important in medical applications, and the measurement and characterization based on fluorescence analysis can be regarded as an important basis for DNA research. Therefore, the use of new fluorescent labeling technology, especially the use of quantum dots to label DNA is a new development in the recent res...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 谢海燕章睿赵东旭
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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