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Electrochemical logic operation method based on DNA template-free amplification and metal ion-glutathione switch

A technology of glutathione and metal ions, applied in the field of logic gate operation, to achieve the effect of rapid analysis, high catalytic activity and high flexibility

Active Publication Date: 2020-03-20
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no electrochemical logic operation method combining DNA template-free amplification and metal ion-glutathione switch at home and abroad

Method used

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  • Electrochemical logic operation method based on DNA template-free amplification and metal ion-glutathione switch
  • Electrochemical logic operation method based on DNA template-free amplification and metal ion-glutathione switch
  • Electrochemical logic operation method based on DNA template-free amplification and metal ion-glutathione switch

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The preparation of embodiment 1 sensor

[0025] (1) Gold electrode (Au): Al with a diameter of 0.3 μm is used before the gold electrode is used 2 o 3 The powder was ground, ultrasonically cleaned 3 times with ultrapure water, dried in a nitrogen stream, and soaked in H 2 SO 4(1.0M) solution, carry out cyclic voltammetry scanning in the range of -0.4V ~ +1.3V for 10min, and finally wash with ultrapure water and blow dry with nitrogen gas for later use.

[0026] (2) Electrode 1: Take a thiol DNA solution (1.0 μM, 5 μL) and drop-coat it on the surface of Au, incubate overnight in a refrigerator at 4°C, wash the electrode slowly with distilled water, treat it with 1.0 mM mercaptohexanol (MCH) for 30 min, and slowly wash it with distilled water. Buffer wash.

[0027] (3) Electrode 2: Mix 1 μL distilled water, 1 μL TdT buffer, dNTP (0.4 μL dTTP, 0.6 μL dGTP, 10 mM), TdT (0.5 μL, 100 U / mL), and drop them on the surface of Electrode 1, at 37 ° C After standing for 1 h, the...

Embodiment 2

[0030] Embodiment 2 Feasibility experiment

[0031] The process of preparing the sensor is the same as in Example 1, Electrode 3 is placed in PBS solution (containing 2mM TMB, 2mM H 2 o 2 ) to scan the response of the electrode to the solution by cyclic voltammetry, and then the electrode was taken out and placed in 1mM Ag + Incubate in the solution for 30min, place in PBS solution (containing 2mM TMB, 2mM H 2 o 2 ) using cyclic voltammetry to scan the response of the electrode to the solution; then the electrode was taken out and placed in 1mM GSH solution for incubation for 30min, then placed in PBS solution (containing 2mM TMB, 2mMH 2 o 2 ), scan the response of the electrode to the solution by cyclic voltammetry, and repeat the operation. The result is as figure 2 As shown, Ag + The addition of GSH will lead to the disappearance of the electrochemical signal (G4 is destroyed), and the addition of GSH will lead to the restoration of the electrochemical signal, thus ...

Embodiment 3

[0032] Example 3 Construction of logic gates

[0033] As shown in Table 1, in the NOT logic gate, for "input", "1" means Ag + join, "0" means Ag + Not joined. For "Output" "1" means the signal is generated, "0" means the signal is not generated, Hg 2+ Same as Ag + design.

[0034] Table 1

[0035] Input (Ag + )

[0036] As shown in Table 2, in the IMPLY logic gate, for "input", "1" means Ag + or GSH added, "0" means Ag + Or GSH is not added. For "Output" "1" means the signal is generated, "0" means the signal is not generated, Hg 2+ -GSH same as Ag + -GSH design.

[0037] Table 2

[0038] Input (Ag + )

[0039] As shown in Table 3, in the NOR logic gate, for "input", "1" means Ag + or Hg 2+ join, "0" means Ag + or Hg 2+ Not joined. For "Output", "1" indicates that the signal is generated, and "0" indicates that the signal is not generated.

[0040] table 3

[0041] Input (Ag + )

[0042] The results in Table 1, Table 2, and...

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Abstract

The invention discloses a series of electrochemical logic operation methods constructed on the basis of DNA template-free amplification and a metal ion-glutathione switch, and the method comprises thefollowing specific steps: pretreating a gold electrode (Au), dropwise coating a DNA solution on the surface of the Au, incubating in a refrigerator at 4 DEG C overnight, and treating with mercaptohexanol (MCH); uniformly mixing distilled water, a TdT buffer solution, dNTP (dTTP+dGTP) and TdT, and dripping the mixture on the surface of the electrode; after the polymerization reaction, adding a buffer solution (Tris-HCl, KCl, pH 7.4), and incubating at room temperature; dropwise adding hemin to the electrode, and incubating at room temperature; after the electrode is prepared in each step, slowly washing the electrode with distilled water to remove impurities. Two NOT logic gates, two IMPLY logic gates and one NOR logic gate are constructed. The method has the advantages of simple preparation, high detection speed, accurate and reliable result and low cost.

Description

technical field [0001] The invention relates to a logic gate operation method, in particular to an electrochemical logic operation method based on DNA template-free amplification and metal ion-glutathione switch, and belongs to the technical field of functional biological materials and biosensing. Background technique [0002] Logic gates are the basic components of integrated circuits. Simple logic gates generally consist of transistors. The high and low levels representing the two signals can be combined by transistors in the integrated circuit to generate a high or low level signal. These two signals can be represented by "true" and "false" or 1 and 0 in binary, respectively. With the rapid development of biotechnology, logic gates have been gradually applied in the field of biotechnology. DNA has the characteristics of sequence-specific recognition, and is easy to synthesize and has good biocompatibility. It has been used to construct molecular logic gates in vitro an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/48
CPCG01N27/3275G01N27/48
Inventor 胡宇芳张青青胡丹丹詹甜玉王邃郭智勇
Owner NINGBO UNIV
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