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Gold electrode modified with specific dna pseudoknot structure, preparation method and application

A gold electrode and specific technology, which is applied in the gold electrode and preparation field of specific DNA pseudoknot structure modification, can solve the problems of complex DNA chain design and low binding efficiency, and achieve high specificity, good repeatability, and simple operation Effect

Active Publication Date: 2018-04-10
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods still have the disadvantages of complicated DNA chain design and low binding efficiency.

Method used

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  • Gold electrode modified with specific dna pseudoknot structure, preparation method and application
  • Gold electrode modified with specific dna pseudoknot structure, preparation method and application
  • Gold electrode modified with specific dna pseudoknot structure, preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Using a gold electrode modified with a specific DNA pseudoknot structure to detect different concentrations of Nanog.

[0021] Step 1, soak the gold electrode (3.0mm diameter) in the piranha solution (H 2 SO 4 :30%H 2 o 2 =3:1) for 15 minutes to eliminate adsorbed substances and rinse with ultrapure water. Then, they were polished with 1 μm and 0.3 μm alumina, respectively, and ultrasonicated in ethanol and ultrapure water for 10 minutes, and dried with nitrogen. Then the piranha solution was added dropwise to the surface of the gold electrode to react for 5 minutes, rinsed with ultrapure water, and dried.

[0022] Step 2, place the treated gold electrode in 0.5M H 2 SO 4 In the process, voltammetric scanning is carried out in the voltage range of 0-1.6V, and the scanning speed is 100mV / s until it reaches a stable state; after the gold electrode is rinsed with ultrapure water and dried, it is immersed in the electrode modification solution containing DN...

Embodiment 2

[0025] Example 2: Biosensor specificity research.

[0026] In order to verify the specificity of the present invention to detect Nanog, the present invention uses different proteins, such as human hemoglobin, bovine serum albumin (BSA) and transcription factor NF-κB, to verify the biosensor's biosensor according to the operation process of Example 1 specificity. Such as image 3 As shown, when there is no Nanog or replaced by other non-specific proteins, there is almost no peak, only when Nanog is added, an obvious peak signal is detected. Experiments have confirmed that this method has good specificity, and the detection of this biosensor has high selectivity.

[0027]

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Abstract

The invention specifically relates to a specific DNA pseudoknot structure-modified gold electrode, and a preparation method and application thereof, belonging to the technical field of electrochemical sensors. The surface of the gold electrode is modified by a self-assembled DNA pseudoknot structure in virtue of action of a gold-mercapto bond, wherein the terminal of the DNA pseudoknot structure has a methylene blue redox label and the stem-loop 1 of the DNA pseudoknot structure contains a Nanog binding sequence. A biosensor utilizes the functionalied DNA pseudoknot structure probe-modified gold electrode for reaction with target protein, which leads to changes of the conformation of the DNA pseudoknot structure; and in virtue of an electric signal molecule MB with good conductivity, sensitive detection of the transcription factor Nanog is realized, and target protein can be effectively distinguished from other control proteins. A detection method provided by the invention is simple to operate and has high sensitivity and specificity and a minimum detection range of 170 pM.

Description

technical field [0001] The invention belongs to the technical field of electrochemical sensors, and in particular relates to a gold electrode modified with a specific DNA pseudoknot structure, a preparation method and an application thereof. Background technique [0002] One of the most intriguing questions in cell biology is cell pluripotency, which is tightly regulated and whose dysregulation can lead to birth defects, failure to repair damage, and even cancer. Nanog is one of the important transcription factors (transcription factors, TFs) related to pluripotency. It is a biomolecule that has attracted great interest recently. Its function is to determine cell fate by regulating cell proliferation and death. It is known that Nanog is not expressed in most normal adult tissues, but is highly expressed in embryonic stem cells and tumor cells. Studies have revealed that Nanog contributes to the stem cell-like phenotype of tumor cells in tumors. Elevated Nanog levels are of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327G01N27/48
CPCG01N27/3277G01N27/48
Inventor 李根喜马洁桦李超陶雅沁
Owner NANJING UNIV