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Method for detecting cancer marker MicroRNA based on three-dimensional graphene biosensor

A biosensor and graphene technology, applied in biochemical equipment and methods, microbial measurement/inspection, instruments, etc., can solve the problems of limited detection range, complicated steps, low sensitivity, etc., and achieve simple transfer method, simple operation, highly sensitive effect

Pending Publication Date: 2020-01-17
DEZHOU UNIV
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  • Application Information

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Problems solved by technology

[0004] At present, traditional miRNA detection techniques mainly include Northern blotting, quantitative real-time polymerase chain reaction (qRT-PCR), microarray and other methods. However, these methods still have many shortcomings, such as time-consuming operation, complicated steps, and sensitivity low and limited detection range, etc., so it is very urgent to develop a sensitive and efficient miRNA detection method

Method used

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  • Method for detecting cancer marker MicroRNA based on three-dimensional graphene biosensor
  • Method for detecting cancer marker MicroRNA based on three-dimensional graphene biosensor
  • Method for detecting cancer marker MicroRNA based on three-dimensional graphene biosensor

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Embodiment 1 3

[0080] The preparation of embodiment 1 three-dimensional graphene biosensor

[0081] Such as figure 1 As shown, the manufacture of a three-dimensional graphene biosensor device needs to go through the process of etching the substrate, transferring, and pasting the sample cell.

[0082] A preparation method of a three-dimensional graphene biosensor device, comprising the following steps:

[0083] (1) Glass covered with indium tin oxide (ITO) conductive film is selected as the substrate. Indium tin oxide is arranged on both sides of the glass substrate, one side is the source and the other side is the drain. The source and drain resistances are both 1KΩ. The size of the glass substrate is 30×30 mm, the size of the indium tin oxide conductive film is 30×12 mm, and the thickness is 185 nm. The substrate was ultrasonically cleaned with acetone, ethanol and deionized water in sequence, and the cleaning time was 20 min each time to make the surface of the substrate cleaner.

[0...

Embodiment 2

[0089] The detection of embodiment 2 complementary miRNA

[0090] The method for detecting miRNA based on a three-dimensional graphene biosensor, the steps are as follows:

[0091] (1) Put the three-dimensional graphene biosensor device on the probe station and connect it to the detection circuit;

[0092] (2) Add 0.1x phosphate buffer saline (PBS, pH 7.0) to test the transmission characteristics of the empty device, the results are as follows figure 2 As shown, and the gate voltage range of the adjustment detection circuit is -1V~1V, and the constant voltage of the source-drain electrode in the adjustment detection circuit is 0.5V.

[0093] (3) Aspirate the PBS out of the sample pool, wash it with 0.1xPBS, add 100mM 1-pyrenebutyric acid N-hydroxysuccinimide ester (PBASE) and incubate for 1h, measure the transmission characteristics, and the test results are as follows: image 3 shown.

[0094] (4) Aspirate PBASE out of the sample cell, wash it with 0.1xPBS, dissolve the p...

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Abstract

The invention relates to a method for detecting a cancer marker MicroRNA based on a three-dimensional graphene biosensor. The three-dimensional graphene biosensor consists of a glass substrate and a three-dimensional graphene layer. Two sides of the upper surface of the glass substrate are provided with indium tin oxide (ITO), and the upper surface of part of indium tin oxide on both sides are covered with the three-dimensional graphene layer. The part uncovered with the three-dimensional graphene on one side serves as a source electrode, and the part uncovered with the three-dimensional graphene on the other side serves as a drain electrode. The method of the invention requires no labeling, is simple to operate and convenient to use, has higher selectivity and specificity under lower usevoltage, and has good safety.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid detection, and in particular relates to a method for detecting cancer marker MicroRNA based on a three-dimensional graphene biosensor. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] MicroRNA (miRNA) is a class of non-coding single-stranded RNA molecules with a length of about 18-24 nucleotides, mainly encoded by endogenous genes, which can regulate the expression of eukaryotic genes and participate in various biological processes, such as The processes of cell proliferation, differentiation, apoptosis, RNA processing and translation play a key role in biological gene regulation, which can...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6825G01N27/414
CPCC12Q1/6825G01N27/4145C12Q2525/207C12Q2565/607
Inventor 许士才宋瑞洪李崇辉王吉华田蒙李迎仙
Owner DEZHOU UNIV
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