A kind of electrochemical biosensor and its preparation method based on three-strand linked DNA
A biosensor and electrochemical technology, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., can solve the problems of high cost, time-consuming, poor repeatability, etc., achieve high versatility, low background signal, and reduce false positives The effect of probability
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Embodiment 1
[0046] Example 1, an ultrasensitive electrochemical biosensor based on triple-stranded DNA was used to detect different concentrations of target miRNA-21 in solution, and a standard working curve of the sensor's current response was obtained.
[0047] (1) A gold electrode with a diameter of 2mm is covered with Al 2 o 3 Grinding with polishing powder, rinsed with ultrapure water;
[0048] (2) Take 10µL, 1µM capture probe immobilization solution and drop it on the electrode surface, cover it, incubate at room temperature for 180min, wash the electrode surface with buffer solution, N 2 blow dry;
[0049] (3) Take 10µL, 2mM mercaptohexanol solution and drop it on the surface of the electrode obtained in step (2), cover it, incubate at room temperature for 90min, seal the non-specific active sites on the electrode surface, wash the electrode surface with buffer solution, N 2 blow dry;
[0050] (4) Take 10µL of hybridization buffer containing different concentrations of target p...
Embodiment 2
[0069] Example 2, the ultrasensitive electrochemical biosensor based on triple-stranded concatenated DNA is applied to the detection of miRNA-21 content in actual serum samples
[0070] (1) Construct an electrochemical biosensor according to the preparation method of the present invention, and use an electrochemical workstation for testing with a three-electrode system, silver / silver chloride is used as a reference electrode, a platinum wire electrode is used as a counter electrode, and the prepared sensor is used as a working electrode , test in 8ml, 10mM pH 6 electrochemical detection solution (containing serum sample, the volume ratio of serum sample to 10mM electrochemical detection solution is 1:1);
[0071] (2) Use differential pulse voltammetry to detect the above electrochemical detection solution, the potential range is -0.6~0.2, the amplitude is 0.05V, the pulse width is 0.05s, and the pulse period is 0.5s;
[0072] (3) As shown in Figure 4, the expression of miRNA-2...
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