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Lead ion detection equipment based on DNA hydrogel and preparation and detection methods thereof

A detection method, hydrogel technology, applied in the field of pollutant detection, can solve the problems that cannot meet the needs of trace lead ion detection, lack of signal amplification strategy, high detection limit, etc., to achieve accurate detection and increase detection sensitivity , the effect of easy operation

Inactive Publication Date: 2020-06-05
CHONGQING UNIV OF ARTS & SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But the detection means of prior art needs to use the labeled nucleic acid of fluorophore and quenching group, the sensitivity of detection is limited; 2+ The minimum detection limit is 20nM), can not meet the needs of trace lead ion detection

Method used

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  • Lead ion detection equipment based on DNA hydrogel and preparation and detection methods thereof
  • Lead ion detection equipment based on DNA hydrogel and preparation and detection methods thereof
  • Lead ion detection equipment based on DNA hydrogel and preparation and detection methods thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] 1. Preparation of the complex:

[0048] Take 5 μL of the capture chain solution with a concentration of 100 μM, add 195 μL of Tirs-HCl (pH=8) buffer solution for dilution, shake well, prepare a capture chain solution with a concentration of 2.5 μM, and store it in a refrigerator at -20°C until use. Then, 0.05 g of acrylamide was accurately weighed, and 0.95 mL of Tirs-HCl buffer solution was added to prepare an acrylamide solution with a mass fraction of 5%. Then accurately pipette 4 μL (100 μM) of the substrate chain solution and 16 μL of 5% acrylamide solution, mix and shake well (mixed solution A), and simultaneously pipette 4 μL (100 μM) of the enzyme chain solution and 16 μL of 5% acrylamide solution Mix and shake well (mixed solution B). Then the mixed solutions (mixed solutions A and B) were placed in a vacuum oven at 37° C. for 10 min. Freshly prepared 0.28 μL of APS (ammonium persulfate) (10%, w / w APS) and 0.56 μL of TEMED (N,N,N',N'-tetramethylethylenediamin...

Embodiment 2

[0061] In this example, the detection system established in Example 1 is used to detect the samples. In order to verify the Pb of this scheme 2+ Whether the detection method is practical in actual sample analysis, we add different concentrations of Pb to different samples (including tap water and lake water) 2+ solution to obtain the sample to be tested (the situation of the sample to be tested is as shown in Table 1, "the added Pb 2+ Concentration" indicates the actual Pb in the sample 2+ concentration), and use the detection system constructed in Example 1 (including the standard curve), to carry out the determination of EIS for the sample to be tested and the electrode to be tested. The specific operation is as follows: according to the method of Example 1, the glassy carbon electrode covered with gold nanoparticles was covered with DNA hydrogel, and then 8 μL of the sample to be tested was dropped on the modified electrode, and incubated in an oven at 37 ° C for 1 h. Af...

experiment example 1

[0066] Experimental example 1: Optimization of the incubation time of DNA hydrogel

[0067] Optimization of DNA hydrogel incubation time is based on Pb 2+ The length of the incubation time leads to the change of the electrochemical impedance value to determine, using different incubation times to detect the impedance value (the DNA hydrogel solution wrapped with protein molecules is added to the electrode surface, and then incubated). The experimental results are as follows Image 6 shown, from Image 6It can be seen that the impedance value gradually increases with the extension of the incubation time. This is because the longer the incubation time, the more DNA hydrogel is formed on the surface of the electrode. Using the three-dimensional network structure of the DNA hydrogel, BSA is wrapped in the hydrogel, and the hydrogel and BSA have a strong effect on ferricyanide ions. The greater the resistance to diffusion, the greater the resistance value. After 120 min, the imp...

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Abstract

The invention belongs to the field of pollutant detection, and relates to lead ion detection equipment and a preparation and detection method thereof, in particular to lead ion detection equipment based on DNA hydrogel and a preparation and detection method thereof. According to the scheme, an electrochemical biosensor is constructed by adopting a signal amplification strategy that protein is wrapped by DNA hydrogel formed by assistance of deoxyribozyme of which the lead ion is dependent, then lead ions are detected, and the method has the advantages of being good in linear range, low in detection limit, high in accuracy, good in specificity and the like. The equipment and the method can be applied to detection practice of lead ions and development and research of novel detection equipment.

Description

technical field [0001] The invention belongs to the field of pollutant detection, and relates to a lead ion detection device and a preparation and detection method thereof, in particular to a DNA hydrogel-based lead ion detection device and a preparation and detection method thereof. Background technique [0002] The heavy metal industry has played an important role in promoting my country's social and economic development and scientific and technological progress. With the increasing utilization of heavy metal products, people have gradually realized that the damage of heavy metal elements to the ecological environment is very serious and threatens the survival and safety of human beings. As we all know, lead is one of the heavy metals, due to the lead ion (Pb 2+ ) cannot be biodegraded in the environment, and can be enriched in organisms through the food chain, and can even be transformed into more toxic chemical forms. Therefore, after heavy metal ions enter the organism...

Claims

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Application Information

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IPC IPC(8): G01N27/02G01N27/327C12Q1/00
CPCC12Q1/002C12Q1/005G01N27/021G01N27/3275G01N2333/922
Inventor 谢顺碧彭琴滕柳梅唐英张进
Owner CHONGQING UNIV OF ARTS & SCI
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