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Photoelectric chemical biosensor as well as preparation method and application of photoelectric chemical biosensor

A biosensor, photoelectrochemical technology, applied in the field of sensors, can solve the problems of limited photoelectric signal and sensitivity of photoelectric sensors, and achieve the effect of improving detection sensitivity

Active Publication Date: 2020-09-22
QINGDAO UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, at present, the photoelectric signal and sensitivity of photoelectric sensors are limited. After a lot of research, it has been found that manganese porphyrin is a metal complex that integrates manganese into porphyrin, which has low production cost, stable chemical performance, high catalytic performance and superior biological Compatibility and other advantages, therefore, can be used in photoelectrochemical analysis

Method used

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  • Photoelectric chemical biosensor as well as preparation method and application of photoelectric chemical biosensor
  • Photoelectric chemical biosensor as well as preparation method and application of photoelectric chemical biosensor
  • Photoelectric chemical biosensor as well as preparation method and application of photoelectric chemical biosensor

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Effect test

Embodiment 1

[0055] Mix 1 μL of padlock DNA with a concentration of 100 μM, 3 μL of assistantDNA with a concentration of 100 μM, 4 μL of secondary water and 3 μL of a buffer solution of 10×T4 DNA ligase, and anneal at 95°C for 5 min, then naturally cool to room temperature, and then add 120 U of T4DNA ligase was added to the mixed solution and connected at 37°C for 1h, then the mixed solution was reacted at 65°C for 10min to inactivate T4DNA ligase, after cooling to room temperature, add 4μL exonuclease buffer solution, 20U exonuclease I and 100U exonuclease III, cutting at 37°C for 2h, then reacting the mixed solution at 80°C for 15min to inactivate exonuclease I and exonuclease III, after cooling to room temperature, the DNA rolling circle template can be obtained , and store the obtained DNA rolling circle template at 4°C for future use.

[0056] Anneal 18 μL of HP with a concentration of 1 μM at 95°C for 5 minutes, then cool naturally to room temperature to obtain the hairpin structure...

Embodiment 2

[0059] Mix 1 μL of padlock DNA with a concentration of 100 μM, 3 μL of assistantDNA with a concentration of 100 μM, 4 μL of secondary water and 3 μL of a buffer solution of 10×T4 DNA ligase, and anneal at 97°C for 4 min, then naturally cool to room temperature, and then add 130 U of T4DNA ligase was added to the mixed solution and connected at 37°C for 2h, then the mixed solution was reacted at 68°C for 8min to inactivate T4DNA ligase, after cooling to room temperature, add 4μL exonuclease buffer solution, 30U exonuclease I and 110U exonuclease III, cutting at 37°C for 2h, then reacting the mixed solution at 83°C for 12min to inactivate exonuclease I and exonuclease III, after cooling to room temperature, the DNA rolling circle template can be obtained , and store the obtained DNA rolling circle template at 6°C for future use.

[0060] Anneal 18 μL of HP with a concentration of 1 μM at 97°C for 4 minutes, then cool naturally to room temperature to obtain the hairpin structure,...

Embodiment 3

[0063] Mix 1 μL of padlock DNA with a concentration of 100 μM, 3 μL of assistantDNA with a concentration of 100 μM, 4 μL of secondary water and 3 μL of a buffer solution of 10×T4 DNA ligase, and anneal at 93 °C for 6 min, then naturally cool to room temperature, and then add 1110 U of T4DNA ligase was added to the mixed solution and connected at 35°C for 1h, then the mixed solution was reacted at 62°C for 12min to inactivate T4DNA ligase, after cooling to room temperature, add 4μL exonuclease buffer solution, 10U exonuclease I and 90U exonuclease III, cutting at 35°C for 1h, then reacting the mixed solution at 77°C for 18min to inactivate exonuclease I and exonuclease III, after cooling to room temperature, the DNA rolling circle template can be obtained , and store the obtained DNA rolling circle template at 2°C for future use.

[0064] Anneal 18 μL of HP with a concentration of 1 μM at 93°C for 6 minutes, then cool naturally to room temperature to obtain the hairpin structur...

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Abstract

The present invention discloses a photoelectric chemical biosensor based on a DNA nanometer dendrite amplifying signal as well as a preparation method and application of the photoelectric chemical biosensor. A technical solution of the present invention is that trace DNA triggers an exonuclease III to provide assistance for a cyclic amplification reaction so as to generate a great number of product chains, DNA rolling circle amplification is combined with chained hybridization to form a nanometer dendrite, a great deal of manganese porphyrin is embedded into the nanometer dendrite to quench abismuth sulfide photoelectric signal, and thus, a simple, convenient and practical photoelectric chemical (PEC) sensor is prepared to realize ultrasensitive detection of the trace DNA and achieve a favorable application potential in fields such as clinical diagnosis, gene therapy, environmental monitoring and food safety.

Description

technical field [0001] The invention relates to the technical field of sensors, and more specifically relates to a photoelectrochemical biosensor and its preparation method and application. Background technique [0002] Currently, there is an increasing need for rapid, highly specific methods for the detection and quantification of chemical, biochemical, and biological substances. Most urgently needed are methods to measure small quantities of drugs, metabolites, microbes, and other substances of diagnostic value. Such as anesthetics, poisons, drugs used for therapeutic purposes, hormones, pathogenic microorganisms, viruses, antibodies, metabolites, enzymes and nucleic acids, etc., the detection of specific trace DNA sequences is useful in clinical diagnosis, gene therapy, environmental monitoring, food safety, etc. Fields are very critical. [0003] Photoelectrochemical (PEC) biosensing is a new detection method developed by combining photoelectrochemical analysis technol...

Claims

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Application Information

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IPC IPC(8): C12Q1/682C12Q1/6825G01N27/26G01N27/327
CPCC12Q1/682C12Q1/6825G01N27/26G01N27/3278C12Q2531/125C12Q2521/501C12Q2521/319C12Q2565/607C12Q2525/301C12Q2563/137
Inventor 接贵霞接贵芬
Owner QINGDAO UNIV OF SCI & TECH
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