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Fluorescence detection method of potassium ion

A fluorescence detection and potassium ion technology, applied in the field of fluorescence detection of potassium ions, can solve the problems of low sensitivity and selectivity, limit practical application, and high detection cost, and achieve the effect of simplifying operation steps, saving detection time, and high sensitivity

Inactive Publication Date: 2012-10-03
LUDONG UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, methods for detecting potassium ions based on conformational changes of nucleic acid aptamers have been reported [UEYAMA H, TAKAGI M, TAKENAKA S.A novel potassium sensing in aqueous media with a synthetic oligonucleotide derivative. Fluorescence resonance energy transfer associated with guanine quartet-potassium ion complex formation[J].J Am Chem Soc,2002,124(48):14286-14287.HO H A, LECLERC M.Optical sensors based on hybrid aptamer / conjugated polymer complexes[J].J Am Chem Soc,2004,126( 5): 1384-1387.], but there are still shortcomings of low sensitivity and selectivity, and some need to carry out DNA double labeling, and the detection cost is high, which limits its practical application
Recently Choi et al [CHOI M S, YOON M, BAEG J O, KIM J.Label-free dual assay of DNA sequences and potassium ions using an aptamer probe and a molecular light switch complex[J].Chem Commun, 2009: 7419-7421. ]Using ruthenium complexes and nucleic acid aptamers to achieve double detection of DNA sequences and potassium ions, but the method needs to be reacted at 70°C and needs to be cooled in an ice-water bath, and the operation is cumbersome

Method used

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  • Fluorescence detection method of potassium ion
  • Fluorescence detection method of potassium ion
  • Fluorescence detection method of potassium ion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Add 2mL buffer (50mM Tris-HCl, pH 7.40) to the cuvette, add 5μL 4×10 -5 Mthiazole orange, 5 μL 6 × 10 -5 M Potassium ion-specific DNA and 5 μL 0.4M potassium chloride were mixed evenly and reacted at 25°C for 4 minutes, and the fluorescence test was performed. At this time, the final concentration of each substance was: [thiazole orange]=100 nM, [potassium ion-specific DNA]=150 nM, [potassium ion]=1 mM.

[0030] Fluorescence spectra before and after adding potassium ions figure 1 As shown, before the addition of potassium ions, the fluorescence intensity at the maximum emission wavelength is 353.06, and after the addition of potassium ions, the fluorescence intensity at the maximum emission wavelength is 151.42, so the present invention can quantitatively detect potassium ions.

Embodiment 2

[0032] Potassium ions are replaced by other metal cations such as calcium ions, magnesium ions, lithium ions, ammonium ions, and sodium ions repeat the steps of Example 1, and the results are as follows figure 1 shown. Calcium ions, magnesium ions, lithium ions, ammonium ions, and sodium ions can also reduce the fluorescence intensity, but compared with potassium ions, the reduction is very small. It shows that the present invention is less interfered by other metal cations and has better selectivity.

Embodiment 3

[0034] Change the potassium ion concentration into the potassium ion of different concentrations and repeat the steps of Example 1, the results are as follows figure 2 shown. It shows that the present invention can quantitatively detect potassium ions of different concentrations. When [thiazole orange]=100nM, [potassium ion-specific DNA]=150nM, the concentration of potassium ion is proportional to the fluorescence intensity in the range of 1-60μM and 400-900μM.

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Abstract

The invention discloses a fluorescence detection method of a potassium ion. The specific DNA (5'-GGGTTAGGGTTAGGG TTAGGG-3') of the potassium ion is in an irregular curling state when no potassium ion exists, fluorescent dye thiazole orange and a G-rich sequence are acted to generate very strong fluorescence, after the potassium ion is added, the specific DNA of the potassium ion forms a G tetrahedral structure, and the fluorescence is decreased. The fluorescence detection method disclosed by the invention not only has high sensitivity and better specificity, but also is simple and rapid, and the whole process can be completed in five minutes.

Description

technical field [0001] The invention relates to the detection field of potassium ions, in particular to a fluorescence detection method for potassium ions. Background technique [0002] Potassium is an indispensable constant element in the human body, and its main function is to maintain the normal function of nerves and muscles. K + It can participate in biological processes such as nerve impulse transmission and cell activity regulation. The content of potassium ions is an important indicator of human physiological activities, and the level in blood and urine is particularly important for clinical diagnosis [KOFUJI P, NEWMAN E A. Potassium buffering in the central nervous system [J]. Neuroscience, 2004, 129: 1045-1056.], so the determination of potassium ion content is of great significance. At present, there are many methods for potassium ion detection, such as flame photometry, ion selective electrode method, atomic absorption spectrometry, potentiometric titration, et...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
Inventor 徐慧陈建农高淑丽柳全文
Owner LUDONG UNIVERSITY
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