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
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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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