Method for detecting affinity of guanine riboswitch based on graphene biosensor

A biosensor, riboswitch technology, applied in instruments, measuring devices, scientific instruments, etc., can solve the problems of limited detection range, complicated steps, low sensitivity, etc., and achieve the effects of low sensitivity, simple operation, and high detection sensitivity

Active Publication Date: 2019-02-19
DEZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods are time-consuming, complicated steps, low sensitivity and limited detection range

Method used

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  • Method for detecting affinity of guanine riboswitch based on graphene biosensor
  • Method for detecting affinity of guanine riboswitch based on graphene biosensor
  • Method for detecting affinity of guanine riboswitch based on graphene biosensor

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] The preparation of embodiment 1 graphene biosensor

[0054] Such as figure 1 As shown, the manufacture of a graphene biosensor device needs to go through the processes of uniform glue, heating and curing, etching the substrate, transferring, removing glue, and pasting the sample cell.

[0055] A preparation method of a graphene biosensor device, comprising the following steps:

[0056] (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.

[0057...

Embodiment 2

[0064] Example 2 Detection of Guanine Riboswitch Affinity

[0065] The method for detecting guanine riboswitch affinity based on graphene biosensor, the steps are as follows:

[0066] (1) Place the graphene biosensor device on the probe station and connect to the detection circuit;

[0067] (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.

[0068] (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.

[0069] (4) Aspirate PBASE out of the sample pool, wash it with 0.1xPBS, dissolve the guanine riboswitc...

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Abstract

The invention discloses a method for detecting an affinity of a guanine riboswitch based on a graphene biosensor. The graphene biosensor is composed of a glass substrate and a graphene layer. Indium tin oxide (ITO) is arranged on two sides of the upper surface of the glass substrate. A part of the upper surface of the indium tin oxide on the two sides are covered by graphene layers, a part on oneside that is not covered by graphene is taken as a source electrode, and a part on the other side that is not covered by graphene is taken as a drain electrode. The method does not need to be marked,is simple to operate and convenient to use. High selectivity and specificity are achieved at a low use voltage, and the safety is good.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid detection and relates to a method for detecting the affinity of a guanine riboswitch based on a graphene biosensor. Background technique [0002] The statements herein merely provide background information related to the present invention and may not necessarily constitute prior art. [0003] Riboswitches are a class of RNA structural elements located on specific regions of non-coding mRNAs that can directly bind small molecular metabolites and regulate gene expression through conformational changes. Riboswitches are highly conserved, can fold into complex three-dimensional structures with pockets, and interact with ligands with high specificity and selectivity, making them a target for a new class of nucleic acid drugs and novel antibiotics. Riboswitch detection can provide very useful information for molecular biology research and genetic disease diagnosis, which is also of great significa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
CPCG01N27/3278
Inventor 许士才田蒙王吉华扈国栋宋瑞洪
Owner DEZHOU UNIV
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