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Three-dimensional DNA nano-structure, electrochemical biosensor as well as preparation methods and application thereof

A biosensor and nanostructure technology, applied in the field of electrochemistry, can solve the problems of residual sequence partial background interference nucleic acid degradation and non-specific synthesis, and achieve the effects of increasing electron transfer rate, improving stability and high sensitivity

Inactive Publication Date: 2015-03-04
NORTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Compared with biosensors based on one-dimensional and two-dimensional DNA nanostructures, although one-dimensional and two-dimensional DNA nanostructure DNA sequences have high specificity in recognizing targets, they are still very susceptible to background interference from the remaining sequence parts, nucleic acid Interference such as degradation and non-specific construction, especially for genetic detection in complex biological environments

Method used

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  • Three-dimensional DNA nano-structure, electrochemical biosensor as well as preparation methods and application thereof
  • Three-dimensional DNA nano-structure, electrochemical biosensor as well as preparation methods and application thereof
  • Three-dimensional DNA nano-structure, electrochemical biosensor as well as preparation methods and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0078] This embodiment is the preparation of hexahedral nanostructure:

[0079] Four DNA strands (P1, P2, P3 and P4, the sequences of the four strands are shown in Table 1) with equimolar mass, the concentration of the four strands is 5.0×10 –6 mol L –1 , added to the 2 and 0.1M NaCl in 20.0mM TE (pH 8.0) buffer, mixed evenly, placed in a 95°C water bath for 30 seconds, and then transferred to a 4°C water bath for 30 seconds to obtain a hexahedral DNA nanostructure.

[0080] Such as figure 1 and 2 As shown, each side of the six faces is composed of the 1-9 sequence in Table 1, where the a, b, c, and d sides are the same, corresponding to the 1, 2 sequence in Table 1; the e side corresponds to the The 3 and 3' sequences; the f side corresponds to the 5 and 5' sequences in Table 1; the g side corresponds to the 7 and 7' sequences in Table 1; the h side corresponds to the 8 and 8' sequences in Table 1.

[0081] The obtained DNA nanostructure was characterized by TEM, and the...

Embodiment 2

[0083] This embodiment is the preparation of biosensor:

[0084] (1) Use 0.3 μm and 0.05 μm γ-Al for the gold electrodes 2 o 3 Polish it to a smooth mirror surface, rinse it carefully with deionized water, and place the electrode in H 2 SO 4 / H 2 o 2 (3:1) mixture, ultrasonically washed for 5 minutes, then ultrasonically washed with ethanol and deionized water for 5 minutes each, and finally dried with nitrogen gas for later use.

[0085] (2) if figure 2 As shown, the treated gold electrode was placed in the solution containing the hexahedral DNA nanostructure prepared in Example 1, and reacted for 10 hours under the condition of avoiding light and shaking, and formed between -SH (mercapto) and the gold electrode (Au). Au-S bond, so that the DNA is assembled on the surface of the electrode, and the electrochemical biosensor is obtained after rinsing with buffer solution.

[0086] The morphology of the electrode surface was characterized by AFM, such as Figure 4 As sh...

Embodiment 3

[0088] This embodiment utilizes the electrochemical biosensor that embodiment 2 obtains to detect thrombin, lysozyme:

[0089] figure 2 The three gold electrodes shown below represent respectively: the hexahedral structure DNA modified gold electrode corresponds to when the solution to be tested contains only thrombin, both thrombin and lysozyme, and only lysozyme, the DNA nanostructure on the electrode surface structure Variety.

[0090] Detection 1: Place the electrode prepared in Example 2 in a solution containing 5.0×10 –10 mol L –1 After reacting for a period of time, the electrode was transferred to a blank buffer solution to record its electrochemical signal. The measurement of the electrochemical cyclic voltammetry spectrum was carried out in a PBS solution containing 0.1M, and the sweep rate was 50mV·s –1 , the potential range is –0.2~1.3V. When no substance to be tested is added to the system, two pairs of redox peaks appear in the cyclic voltammetry curve ( F...

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Abstract

The invention relates to a three-dimensional DNA nano-structure, an electrochemical biosensor as well as preparation methods and application thereof. The three-dimensional DNA nano-structure is a hexahedron structure, and the hexahedron structure can be structurally transformed to cause electrochemical signal transformation by being combined with different target molecules. Four vertexes of the bottom surface of the three-dimensional DNA nano-structure unit are immobilized on the surface of a gold electrode through self-assembly action. According to the invention, the DNA nano-structure is assembled on the surface of the electrode to constitute the novel electrochemical biosensor, the DNA with the hexahedron structure has the characteristics of high-specificity target molecule recognition property and high stability, so that the analysis performance of the electrochemical biosensor is improved. The electrochemical biosensor provided by the invention can realize detection of different to-be-detected objects such as thrombin, lysozyme and the like by replacing a DNA chain and is wide in application range.

Description

technical field [0001] The invention belongs to the technical field of electrochemistry, and specifically relates to a three-dimensional DNA nanostructure, an electrochemical biosensor based on the three-dimensional DNA nanostructure, and a preparation method and application thereof. Background technique [0002] DNA electrochemical biosensor is an analytical tool or system that utilizes the excellent self-assembly and recognition capabilities of DNA molecules, and consists of immobilized biologically sensitive material nucleic acids as recognition elements, electrodes and signal amplification devices. Because of its high sensitivity, low cost, and easy miniaturization, it has become one of the most active fields in analytical chemistry research. However, using one-dimensional (single-stranded DNA) or two-dimensional structure (such as hairpin structure) DNA as the recognition element, the uniformity of the sensing interface is difficult to be effectively controlled during t...

Claims

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Application Information

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IPC IPC(8): G01N27/26G01N27/327G01N27/48
Inventor 盛庆林刘江涛张赛武倩聂菲郑建斌
Owner NORTHWEST UNIV
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