A three-dimensional graphene-based proportional signal amplification aptamer sensor and its preparation method and application
A signal amplification and graphene-based technology, applied in instruments, scientific instruments, measuring devices, etc., can solve the problems of high detection cost, poor selectivity, and low sensitivity, and achieve good biocompatibility, improved accuracy, and improved sensitivity Effect
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Embodiment 1
[0075] Embodiment 1, the preparation of gold nano-three-dimensional graphene composite material (Au-RGO)
[0076] (1) ultrasonically disperse 5 mg of graphene oxide in 1 ml of ultrapure water for 2 hours to obtain a uniformly dispersed graphene oxide suspoemulsion;
[0077] (2) Dilute the graphene oxide suspoemulsion in (1) to 6 mL, then add 200 μL tetrachloroauric acid solution (1%, w / w) and mix well; then add 25 mg polyethylene glycol to 4 mL ultrapure water, After it is fully dissolved, it is added into the mixed solution of graphene oxide and tetrachloroauric acid, and ultrasonically treated for 1 hour to make it fully mixed.
[0078] (3) The mixed solution in (2) was transferred to a 25mL hydrothermal reaction device, reacted in an oven at 160°C for 12h, and then cooled to room temperature. After the reaction product was taken out, it was centrifuged and washed three times with ultrapure water, and the obtained black precipitate was retained.
[0079] (4) Add 8 mL of ul...
Embodiment 2
[0081] Embodiment 2, the preparation of gold nano-three-dimensional graphene composite material (Au-RGO)
[0082] (1) Ultrasonic dispersion of 4 mg graphene oxide in 1 ml ultrapure water for 1 h to obtain a uniformly dispersed graphene oxide suspoemulsion;
[0083] (2) Dilute the graphene oxide suspoemulsion in (1) to 8 mL, then add 150 μL tetrachloroauric acid solution (1%, w / w) and mix well; then add 20 mg polyethylene glycol to 2 mL ultrapure water, After it is fully dissolved, it is added to the mixed solution of graphene oxide and tetrachloroauric acid, and ultrasonically treated for 0.5 h to make it fully mixed.
[0084] (3) Transfer the mixed solution in (2) into a 25mL hydrothermal reaction device, react in an oven at 170°C for 15h, and then cool to room temperature. After the reaction product was taken out, it was centrifuged and washed three times with ultrapure water, and the obtained black precipitate was retained.
[0085] (4) Add 5 mL of ultrapure water to the pr...
Embodiment 3
[0086] Example 3, preparation of gold nanoparticles and methylene blue-labeled MUC1 aptamer-gold nanocomposites:
[0087] (1) get 80mL concentration and be that the tetrachloroauric acid solution of 1mM is in the heating reflux device, be heated to vigorous boiling under the magnetic sub-stirring condition;
[0088] (2) Rapidly add 10 mL of trisodium citrate solution with a concentration of 38.8 mM to (1), and then heat to reflux for 10 min;
[0089] (3) After stopping the heating, cool to room temperature with stirring, and transfer the obtained wine-red gold nanometer into a brown reagent bottle and place it in a 4°C refrigerator for later use.
[0090] Such as image 3 As shown, the diameter of the synthesized uniform gold nanoparticles is about 15nm.
[0091] (4) Take 1 mL of the gold nanoparticles prepared in (3), and add 150 μL of 50 μM methylene blue-labeled MUC1 aptamer solution to it, and incubate at 25° C. for 16 h with slight shaking on a shaker;
[0092] (5) Tra...
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