Preparation method and application of SERS biosensor for detecting tumor marker miRNA-141
A technology of biosensors and tumor markers, applied in the field of detection of tumor marker miRNA-141, can solve the problems of undisclosed SERS biosensors
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Embodiment 1
[0040] A method for preparing a SERS biosensor for detecting tumor marker miRNA-141, comprising the following steps:
[0041] (1) Preparation of capture unit
[0042] a. Preparation of Fe 3 o 4 Magnetic nanoparticles solution: 0.8 g FeCl 3 ·6H 2 O and 0.4 g FeCl 2 4H 2 Dissolve O in 70mL of water, pass nitrogen to remove oxygen, stir and mix evenly, raise the temperature to 90 ℃, slowly add 20-25wt% ammonia water dropwise, adjust pH = 9-10, continue heating and stirring for 1-2 h, stop heating, and Under protection, stir and reflux, cool to room temperature, wash with water until neutral, and dilute to 50 mL with ethanol to obtain Fe 3 o 4 magnetic nanoparticle solution;
[0043] b. Preparation of aminated Fe 3 o 4 Magnetic nanoparticles solution: 45 mL Fe 3 o 4 Sonicate the magnetic nanoparticle solution for 0.7 h, add 0.25 mL of 3-aminopropyltriethoxysilane (APTES), stir for 4-5 h, add 1.5 mL of 0.3 mol / L nitric acid solution, continue stirring for 3-4 h, wash wi...
Embodiment 2
[0057] With above-mentioned embodiment 1, its difference is:
[0058] (1) Preparation of capture unit
[0059] a. Preparation of Fe 3 o 4 Magnetic nanoparticles solution: 0.7 g FeCl 3 ·6H 2 O and 0.2 g FeCl 2 4H 2 O dissolved in 50mL of water;
[0060] b. Preparation of aminated Fe 3 o 4 Magnetic nanoparticles solution: 40 mL Fe 3 o 4 Sonicate the magnetic nanoparticle solution for 0.5 h, add 0.2 mL of 3-aminopropyltriethoxysilane, stir for 4 h, add 1 mL of 0.5 mol / L nitric acid solution, and continue stirring for 3 to 4 h;
[0061] c. Preparation of Fe 3 o 4 @Au magnetic nanoparticle solution: 5 mL of aminated Fe 3 o 4 Magnetic nanoparticles solution and 10 mL of 1wt% HAuCl 4 The solution was mixed at an ultrasonic frequency of 60 kHz, and after stirring for 1 h, 30 mL of 30 mmol / L trisodium citrate solution was slowly added and ultrasonicated for 2 h;
[0062] d. Prepare capture unit solution: Take 50 µL Fe 3 o 4 The @Au magnetic nanoparticle solution was s...
Embodiment 3
[0076] With above-mentioned embodiment 1, its difference is:
[0077] (1) Preparation of capture unit
[0078] a. Preparation of Fe 3 o 4Magnetic nanoparticles solution: 1.0 g FeCl 3 ·6H 2 O and 0.5 g FeCl 2 4H 2 O dissolved in 100mL of water;
[0079] b. Preparation of aminated Fe 3 o 4 Magnetic nanoparticles solution: 50 mL Fe 3 o 4 Sonicate the magnetic nanoparticle solution for 1 h, add 0.3 mL of 3-aminopropyltriethoxysilane, stir for 5 h, add 2 mL of 0.1 mol / L nitric acid solution, and continue stirring for 3 to 4 h;
[0080] c. Preparation of Fe 3 o 4 @Au magnetic nanoparticle solution: 10 mL of aminated Fe 3 o 4 Magnetic nanoparticles solution and 20 mL of 1wt% HAuCl 4 The solution was mixed at an ultrasonic frequency of 100 kHz, and after stirring for 1 h, 50 mL of 20 mmol / L trisodium citrate solution was slowly added, and ultrasonicated for 2 to 3 h;
[0081] d. Prepare capture unit solution: Take 100 µL Fe 3 o 4 The @Au magnetic nanoparticle solutio...
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