DNA aptamer modified bioelectrochemical sensor and preparation method thereof
A DNA aptamer, bioelectrochemical technology, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., can solve problems such as high recurrence rate and low cure rate
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Embodiment 1
[0022] Example 1: Modification of gold electrodes
[0023] Before aptamer modification, the gold electrodes were sanded on fine sandpaper and then on Al 2 o 3 The powder was ground, and then ultrasonicated in absolute ethanol and ultrapure water for 5 minutes. In order to better remove impurities remaining on the surface of the gold electrode, place the electrode in 1M H 2 SO 4 In the 0-1.5V voltage range, the cyclic voltammetry (CV) peak sweep is performed, the sweep speed is set to 100mv / s, and the stability is reached in about 20 cycles. The electrodes were then blown dry in nitrogen for aptamer modification. The gold electrodes were respectively placed on the DNA aptamer chains containing sulfhydryl groups (5'-HS-(CH 2 ) 6 -ATCTA ACTGC TGCGC CGCCG GGAAA ATACT GTACG GTTAG A-3') in Eppendorf tubes containing HS-DNA aptamers in the concentration range of 0.05pM, 0.5pM, 5pM and 50pM, 10mM Tris-HCl, 1mM EDTA, 1M NaCl and 0.5 μM TCEP (pH 8.0), inverted for 16 hours. Afte...
Embodiment 2
[0024] Embodiment two: the cultivation and processing of cells
[0025] CCRF-CEM cells (human acute lymphoblastic leukemia T lymphocytes) and HL-60 cells (human myeloid leukemia cells) were purchased from Shanghai Institute of Cells, Chinese Academy of Sciences. Both cells were maintained at 37 °C, 5% CO 2 The culture medium was 1640 medium containing 10% fetal bovine serum, and the cells were collected when the cells grew to the end of the logarithmic growth phase for cell detection research. Count with a hemocytometer before detection, and the concentration of cells used each time is generally 10 5 about.
Embodiment 3
[0026] Example 3: Optimization of the concentration of the aptamer modified on the gold electrode
[0027] Cyclic voltammetry, the electrochemical probe used is 5mM K 3 [Fe(CN) 6 ] / K 4[Fe(CN) 6 ]. Electrochemical probe detection was performed on the gold electrodes numbered A, B, C, and D above, and the results were as follows: figure 1 . From the results of cyclic voltammetry, it can be observed that with the decreasing concentration of aptamer, K 3 [Fe(CN) 6 ] / K 4 [Fe(CN) 6 ] and the electrochemical transmission of the gold electrode surface becomes easier, so the signal is also increasing continuously, among which the B gold electrode surface and the K 3 [Fe(CN) 6 ] / K 4 [Fe(CN) 6 ] had the best electrochemical transfer, and a better peak shape was obtained. Afterwards, when the aptamer concentration was further reduced, the signal did not increase significantly.
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