Fluorescence analysis method for determining concentration of Hg<2+>

A fluorescence analysis and concentration technology, which is applied in the field of fluorescence analysis to determine Hg2+ concentration, can solve the problems of complicated labeling process, insensitive fluorescence signal, unstable structure, etc., and achieves the effect of low cost, simple and effective operation, and enhanced fluorescence intensity.

Active Publication Date: 2017-05-24
SHANGQIU NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, DNA molecules will form a double-stranded coordination structure under special circumstances, and the DNA molecules used as probes are usually labeled with dye molecules, but the labeling process is complicated and expensive
As another example, G-quadruplex is a kind of DNA with a special structure rich in G bases. It has the characteristic of emitting fluorescence when combined with related substances. Due to its special structure, it can selectively bind to some target molecules Combining with each other produces a change in the fluorescent signal, so it has been used in the design of a variety of biosensors, but most of these designs have problems such as structural instability, insensitivity of the fluorescent signal, and high cost

Method used

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  • Fluorescence analysis method for determining concentration of Hg&lt;2+&gt;
  • Fluorescence analysis method for determining concentration of Hg&lt;2+&gt;
  • Fluorescence analysis method for determining concentration of Hg&lt;2+&gt;

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Embodiment 1

[0026] Example 1 A method for measuring Hg 2+ Concentration Fluorescence Analysis Method

[0027] Select two non-labeled single-stranded DNA molecules A and B with different lengths and rich in T and G bases at the same time, and prepare solution A and solution B with a concentration of 100 μmol / L, respectively. 90°C water bath for 5 min, cooled to room temperature, mixed at a volume ratio of 1:1 to obtain a mixed solution, and set aside; wherein, the nucleotide sequence of the non-labeled single-stranded DNA molecule A is 5'-TTTTTTTGTGGGT-3', As shown in SEQ ID NO: 1; the nucleotide sequence of the non-labeled single-stranded DNA molecule B is 5'-GGGTGGGTGGTTATTTA-3', as shown in SEQ ID NO: 2; non-labeled single-stranded DNA molecules A and B The match length of is 7 T bases.

[0028] Mix 20 μL of the mixture with 10 μL of 200 μmol / L thioflavin solution to obtain mixture I; mix mixture I with 970 μL of 10 mmol / L Tris-HNO 3 The buffer solutions were mixed to obtain the mixe...

Embodiment 2

[0029] Embodiment 2 A method for measuring Hg 2+ Concentration Fluorescence Analysis Method

[0030]Select two non-labeled single-stranded DNA molecules A and B with different lengths and rich in T and G bases at the same time, and prepare solution A and solution B with a concentration of 100 μmol / L, respectively. Water bath at 90°C for 5 min, cooled to room temperature, and mixed at a volume ratio of 1:1 to obtain a mixed solution I for later use; wherein, the nucleotide sequence of the non-labeled single-stranded DNA molecule A is 5'-TTTTTTTTGTGGGT-3' , as shown in SEQ ID NO: 3; the nucleotide sequence of the non-labeled single-stranded DNA molecule B is 5'-GGGTGGGTGGTTATTTAT-3', as shown in SEQ ID NO: 4; the non-labeled single-stranded DNA molecule A and The matching length of B is 8 T bases.

[0031] Mix 20 μL of the mixture with 10 μL of 200 μmol / L thioflavin solution to obtain mixture I; mix mixture I with 970 μL of 10 mmol / L Tris-HNO 3 The buffer solutions were mixed...

Embodiment 3

[0032] Embodiment 3 A method for measuring Hg 2+ Concentration Fluorescence Analysis Method

[0033] Select two non-labeled single-stranded DNA molecules A and B with different lengths and rich in T and G bases at the same time, and prepare solution A and solution B with a concentration of 100 μmol / L, respectively. 90°C water bath for 5 minutes, cooled to room temperature, and mixed at a volume ratio of 1:1 to obtain a mixed solution I, which is set aside; wherein, the nucleotide sequence of the unlabeled single-stranded DNA molecule A is 5'-TTTTTTTTGTGGGT-3', As shown in SEQ ID NO: 5; the nucleotide sequence of the unlabeled single-stranded DNA molecule B is 5'-GGGTGGGTGGTTATTTATT-3', as shown in SEQ ID NO: 6; unlabeled single-stranded DNA molecules A and B The match length of is 9 T bases.

[0034] Mix 20 μL of the mixture with 10 μL of 200 μmol / L thioflavin solution to obtain mixture I; mix mixture I with 970 μL of 10 mmol / L Tris-HNO 3 The buffer solutions were mixed to ...

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Abstract

The invention relates to a fluorescence analysis method for determining the concentration of Hg<2+> and belongs to the technical field of analysis detection. The fluorescence analysis method comprises the following steps: firstly, selecting two non-labeled single-chain DNA (Deoxyribonucleic Acid) molecules with different length and containing rich basic groups including T and G; preparing into solutions respectively; carrying out water-bath treatment for later use; mixing two types of solutions, a thioflavin solution and a Tris-HNO3 buffering solution to obtain a mixed solution; adding the Hg<2+> into the mixed solution, and shaking at a room temperature for 30min; determining a fluorescence intensity value under 450nm; calculating the concentration of the Hg<2+> according to the determined fluorescence intensity value. According to the method provided by the invention, fluorescence is intensively enhanced by self-assembling double-chain / G-quadruplex of the non-labeled DNA molecules induced by the Hg<2+>, and embedding ThT into double-chain DNA and G-quadruplex, so that a complicated DNA fluorescence labeling process is avoided and a fluorescence signal is dually amplified; the fluorescence analysis method is simple to operate, low in cost, good in specificity and high in sensitivity.

Description

technical field [0001] The invention belongs to the technical field of chemical / biological sensing and analysis and detection, and in particular relates to a method for measuring Hg 2+ Concentration fluorescence analysis method. Background technique [0002] Hg 2+ It is a heavy metal ion that is very toxic and cannot be biodegraded. With the continuous development of industry, the mercury in the environment is also increasing. After mercury is absorbed by plants, it will be transferred through the food chain and bio-enrichment. The impact of mercury ions on the environment will gradually be transformed into an impact on human health. When the mercury ion content in the human body accumulates to a certain level, it will lead to the decline of brain, liver and kidney functions, and even death in severe cases. [0003] Traditional detection of Hg 2+ The analysis methods mainly include inductively coupled plasma mass spectrometry (ICP-MS), atomic absorption / fluorescence spec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428
Inventor 王永祥耿凤华姜香宇瞿鹏徐茂田
Owner SHANGQIU NORMAL UNIVERSITY
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