DNA tetrahedron probe for lead ion detection, and lead ion detection method

A tetrahedral probe and lead ion technology, applied in the field of electrochemical detection, can solve the problems of lack of sensitive, simple, accurate and rapid detection of lead ions

Active Publication Date: 2017-12-22
SHANGHAI INST OF MEASUREMENT & TESTING TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The technical problem to be solved by the present invention is to overcome the defects of the lack of sensitive, simple, accurate and fast metho...

Method used

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  • DNA tetrahedron probe for lead ion detection, and lead ion detection method
  • DNA tetrahedron probe for lead ion detection, and lead ion detection method
  • DNA tetrahedron probe for lead ion detection, and lead ion detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] (1) Take 50 μM Tetra-A (its nucleotide sequence is shown in SEQ ID No.1 in the sequence listing), Tetra-B (its nucleotide sequence is shown in SEQ ID No.2 in the sequence listing), Tetra-C (its nucleotide sequence is shown in SEQ ID No.3 in the sequence listing) and Tetra-D (its nucleotide sequence is shown in SEQ ID No.4 in the sequence listing) each 1 μ L, 30mM tris(2- Carboxyethyl) phosphine (TCEP) 1 μL and 45 μL TM buffer solution (composed of 20 mM Tris and 50 mM MgCl 2 composition, pH 8.0) and mix well. After heating at 95°C for 2 minutes, quickly cool down to 4°C for more than 30 seconds, and use a PCR instrument to control the temperature. The DNA tetrahedral nanostructure capture probes containing functionalized nucleic acids were obtained at a final concentration of 1 μM.

[0039] (2) Take the gold plate working electrode (CHI101, purchased from Shanghai Chenhua Company), grind and polish it with 50nm Al2O3 powder for 2min, then use absolute ethanol and Mill...

Embodiment 2

[0049] (3) Mix 100 nM lead ions with 1 μM Pb-deoxyribozyme probe (its nucleotide sequence is shown in SEQ ID No.5 in the sequence listing) in buffer A, and adjust the final volume to 100 μL to obtain a mixed solution a. Buffer A contains 1M NaNO 3 and 20mM MgCl 2 10mM HEPES buffer solution, pH 7.0. Next, the gold electrode assembled with the DNA tetrahedral nanostructure capture probe prepared in step (2) is immersed in the mixed solution A.

[0050] Among them, step (2) prepared 3 groups of gold electrodes assembled with DNA tetrahedral nanostructure capture probes, these 3 groups of gold electrodes were first denatured at 60°C for 5 minutes, then cooled at room temperature for 20 minutes, and then respectively heated at 20°C. After hybridization for 2 hours under the conditions of ℃, 25℃ and 37℃, the hybridized gold electrode was taken out, and the gold electrode was washed with 10 mM HEPES buffer solution.

[0051] All the other steps and parameters are completely consi...

Embodiment 3

[0056] (4) The gold electrode containing the G-quadruple complex structure obtained in step (3) is immersed in a solution containing 1mM H 2 o 2 and Hemin's 10 mM buffer B, followed by electrochemical cyclic voltammetry (CV) testing. Among them, buffer B contains 50mM NaNO 3 and 100 mM KCl in 10 mM HEPES buffer solution, pH 7.0. The concentrations of Hemin were 0.1 μM, 0.2 μM, 0.5 μM, 1 μM, 2 μM and 5 μM, respectively.

[0057] All the other steps and parameters are completely consistent with embodiment 1.

[0058] Detect the current signal, the results are shown in Table 3 and Figure 4 shown. Table 3 shows that the maximum signal-to-noise ratio can be achieved when the concentration of Hemin is 1 μM, so 1 μM is the optimal reaction concentration of Hemin.

[0059] Table 3 The selection of the optimal concentration of Hemin

[0060] Hemin concentration

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Abstract

The present invention provides a DNA tetrahedron probe for lead ion detection, and a lead ion detection method. The DNA tetrahedron probe comprises a single-stranded probe Tetra-A, a single-stranded probe Tetra-B, a single-stranded probe Tetra-C and a single-stranded probe Tetra-D, wherein the 3' terminal of the single-stranded probe Tetra-A contains a structural domain A, and is complementary to a lead ion-dependent deoxyribozyme probe. According to the present invention, the method has the advantages of simple operation and no requirement of expensive instrument of the DNA tetrahedron probe electrochemical detection method, further has the high specificity of the lead ion-dependent deoxyribozyme probe to lead ions, has the lead ion detection range of 10 pM-1000 nM, has advantages of easy performing and reliable result, and can directly detect tap water or pond water and water from other sources.

Description

technical field [0001] The invention belongs to the field of electrochemical detection, and in particular relates to a DNA tetrahedron probe used for the detection of lead ions and a method for detecting lead ions. Background technique [0002] DNA three-dimensional nanostructured probe DNA tetrahedral-structured probe (TSP), which has three thiol-based apexes connected to gold electrodes and a pendant capture probe apex. The study found that the electrochemical sensor using the TSP structure is effective in the detection of cocaine [Anal.Chem., 2011, 83(19): 7418-7423.] and microRNA [Anal.Chem., 2014, 86(5): 2285-2288. ] can better solve the problem of surface crowding effect. [0003] Because lead is widely used in fuels, building materials, coatings, paints, and industrial processes, lead is ubiquitous in the environment, including soil, water, and even the food chain. Lead pollution has always been a very serious environmental problem. Once the inorganic pollutants re...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N27/48
CPCG01N27/327G01N27/3271G01N27/48
Inventor 闻艳丽刘刚王乐乐李兰英杨雪许丽
Owner SHANGHAI INST OF MEASUREMENT & TESTING TECH
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