Preparation method and application of aptamer fluorescence sensor based on aunps@zif-8
A fluorescent sensor, ZIF-8 technology, applied in the field of biosensors, can solve the problems of complex assembly method, complex operation, low sensitivity, etc., and achieve the effects of specificity and sensitivity detection, good biocompatibility, and simple synthesis
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Embodiment 1
[0044] The preparation method of AuNPs@ZIF-8 material includes the following steps:
[0045] (1) Weigh 350 mg of 2-methylimidazole (2-MIM), dissolve in 7 mL of methanol, and dissolve by ultrasonic for 10-15 minutes;
[0046] (2) Weigh 150 mg of zinc nitrate [Zn(NO 3 ) 2 ·6H 2 O] dissolved in 7mL methanol, ultrasonically dissolved for 8-10 minutes;
[0047] (3) under stirring conditions, the zinc nitrate solution obtained in step (2) is slowly added to the 2-methylimidazole solution obtained in step (1), and stirring is continued at room temperature for 20-30 minutes, the obtained product Centrifuged, washed three times with anhydrous methanol, and dried in a vacuum drying oven at 80 °C for 12 h to obtain ZIF-8 material, whose SEM and TEM images are as follows figure 2 As shown in A and 2C, it can be seen from the figure that the ZIF-8 material is a regular dodecahedron structure, the particle size is about 70nm, and the morphology is uniform;
[0048] (4) Weigh 20 mg of ...
Embodiment 2
[0051] A construction process based on AuNPs@ZIF-8 fluorescence sensor, including the following steps:
[0052] (1) Weigh 2 mg of AuNPs@ZIF-8 and dissolve it in 2 mL of Tris-HCl buffer solution (25 mM, pH=7.4), ultrasonicate for about 20 minutes to make the dispersion uniform, and obtain a solution containing AuNPs@ZIF-8;
[0053] (2) Dissolving the purchased DNA sequences DNA S1, DNA Lock1, and DNA DNAzyme in the Tris-HCl buffer solution of 25mM pH=7.4 respectively to obtain DNA S1 solution, DNA Lock1 solution and DNA DNAzyme solution with a concentration of 100uM; The sequences of DNA S1, DNA Lock1, and DNA DNAzyme are as follows:
[0054] DNA S1: 5'-SH-TTTTTTTTTTTTTTGGGCCACTAT rAGGAAT-FAM-3';
[0055] DNA Lock1: 5'-TTGAAG CAC AAA TTC GGT TCT ACA GGG TA-3';
[0056] DNA DNAzymes:
[0057] 5'-SH-TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTATTC TTCCGACCGGTCGAAAATAGTGGCCCGAATTTGTGCTTCAA-3';
[0058] (3) Take 5 μL of the DNA DNAzyme solution obtained in step (2) and add it to ...
Embodiment 3
[0063] Quantitative detection of miRNA-10b by DNAzyme-AuNPs@ZIF-8 fluorescent sensor, the sequence of miRNA-10b is 5'-UACCCUGUAGAACCGAAUUUGUG-3'
[0064] The DNAzyme-AuNPs@ZIF-8 fluorescence sensor solutions obtained in Example 2 with a concentration of 1 mg / mL were divided into multiple groups, and miRNA-10b solutions (which were 2.5OD miRNA-10b solutions) were added to them respectively. 10b was dissolved in DEPC water), incubated at 35 °C for 60 min; then the fluorescence emission spectrum of each system was detected at the excitation wavelength of 490 nm, and the results were as follows Figure 4 as shown in B;
[0065] The standard curve was constructed with the concentration of miRNA-10b solution as the abscissa and the maximum fluorescence intensity of each system as the ordinate. Figure 4 As shown in A, the linear equation is obtained as I=346.26log[c]+936.5728, and its correlation coefficient R 2 The detection limit is 0.9720, and the detection limit is 3 pM. Accor...
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