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One step amplification method of fluorescence detection for mercury ions

A fluorescence detection, mercury ion technology, applied in the field of fluorescence detection, can solve problems such as the inability to form a double-stranded structure, and achieve the effects of fast reaction speed, improved sensitivity, and high-sensitivity detection.

Inactive Publication Date: 2015-11-11
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the absence of mercury ions, the double-chain structure cannot be formed

Method used

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  • One step amplification method of fluorescence detection for mercury ions
  • One step amplification method of fluorescence detection for mercury ions
  • One step amplification method of fluorescence detection for mercury ions

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] (1) Put 7ulH in a 200ul centrifuge tube in sequence 2 O, Probe1 chain 2ul (0.1uM), Probe2 chain 2ul (0.1uM), 1ul target mercury ion, different concentrations of HAP chain 5ul (1uM, 5uM, 10uM, 20uM), Nt.BbvCI buffer 2ul, Nt.BbvCI enzyme 100Uml, make 20ul homogeneous system. Shake on a shaker for 30s and mix well.

[0037] (2) Then put the mixed mixture into a 37°C incubator and incubate for 2h.

[0038] (3) Set the excitation wavelength to 494nm, the emission wavelength range to 500nm-630nm, the incident slit to 5nm, and the exit slit width to 3nm; start testing its fluorescence intensity.

[0039] The experimental results are as follows figure 1 As shown, as the concentration of HAP chains increases, the fluorescence intensity obtained in the experiment increases continuously, until the concentration is 10uM, the fluorescence intensity remains basically unchanged. Therefore, when the HAP chain concentration is 10uM, the experimental effect is the most obvious, and ...

Embodiment 2

[0041] (1) Put 8ulH in a 200ul centrifuge tube in sequence 2 O, Probe1 chain 2ul (0.1uM), Probe2 chain 2ul (0.1uM), 1ul target mercury ion, HAP chain 5ul (50uM), Nt.BbvCI buffer 2ul, Nt.BbvCI enzyme 100Uml, make a 20ul homogeneous system. Shake on a shaker for 30s and mix well.

[0042] (2) Then put the mixed mixture into a 37°C incubator and incubate for different times (30, 60, 90, 120, 150, 180min).

[0043](3) Set the excitation wavelength to 494nm, the emission wavelength range to 500nm-630nm, the incident slit to 5nm, and the exit slit width to 3nm; start testing its fluorescence intensity.

[0044] Under the experimental results figure 2 It shows that with the increase of the incubation time, the fluorescence intensity obtained in the experiment is continuously enhanced, and after the reaction time of 120min, the fluorescence intensity remains basically unchanged. Therefore, when the reaction time is 120min, the experimental raw materials are fully utilized, and the...

Embodiment 3

[0046] (1) In a 200ul centrifuge tube, put 1ul and 8ulH of the target mercury ion in turn 2 O, Probe1 chain 2ul (0.1uM), Probe2 chain 2ul (0.1uM), HAP chain 5ul (10uM), Nt.BbvCI buffer 2ul, add different amount of Nt.BbvCI enzyme (10Uml, 50Uml, 100Uml, 200Uml, 500Uml) Make 20ul homogeneous system. Shake on a shaker for 30s and mix well.

[0047] (2) Then put the mixed mixture into a 37°C incubator and incubate for 120min.

[0048] (3) Set the excitation wavelength to 494nm, the emission wavelength range to 500nm-630nm, the incident slit to 5nm, and the exit slit width to 3nm; start testing its fluorescence intensity.

[0049] The experimental results are as follows image 3 It was shown that with the increase of the amount of endonuclease, the fluorescence intensity obtained in the experiment was continuously enhanced, and after the amount of enzyme reached 100Uml, the fluorescence intensity remained basically unchanged. It shows that the amount of enzyme required to catal...

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Abstract

The invention provides a one step amplification method of fluorescence detection for mercury ions. Detection for the mercury ions is realized on the basis of a principle that the mercury ions can stabilize T-T mismatch in DNA (deoxyribonucleic acid), the specific recognition of a nucleic acid aptamer is used, aptamers of the mercury ions serve as recognition substances to realize the high specific detection for the target mercury ions, reaction speed is improved by using a fluorescence detection method, complexity of the operation is lowered, and fast, simple and flexible detection for the target is realized.

Description

technical field [0001] The invention relates to a method for fluorescence detection of mercury ions by a one-step amplification method, and belongs to the technical field of detection fluorescence. Background technique [0002] Mercury and its compounds have many hazards to human health, and if they exist in natural water bodies, they will pose a threat to a wide range of people. It can accumulate in organisms and transfer to the human body through the biological chain. The trace mercury accumulated in the human body cannot be excreted through its own metabolism, which will directly lead to heart, liver, thyroid disease, and nervous system disorders. , chronic mercury poisoning, and even lead to the formation of malignant tumors. [0003] Dissolved mercury ions often have high chemical activity and are the main form of mercury pollutants discharged into natural water bodies. Their compounds have high water solubility and are also the hub for the transformation of various me...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 王玉黄加栋许颖刘素王虹智郭玉娜崔洁邱婷婷崔雪君裴倩倩冷雪琪韩聪
Owner UNIV OF JINAN
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