A method for rapid release of light-field-regulated genes in gene transfection
A technology for gene transfection and gene regulation, which is applied in the field of biomedicine, can solve the problems of cell activity influence and complicated operation, and achieve the effects of less damage to cells and tissues, simple modification methods, and easy promotion and application
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Embodiment 1
[0023] 1) Diffusion depth of boron atoms is 400nm, diffusion concentration is 1×10 20 atom / cm 2 The p+ / n-type silicon wafers were cut into square pieces of 1cm×1cm and sterilized by high temperature and high pressure for later use. The silicon wafer has good biocompatibility, such as figure 1 shown. The process of preparing the p-DNA / lipofectamine complex is as follows: take an appropriate amount of lipofectamine and disperse it in PBS buffer, and get an appropriate amount of p-DNA and disperse it in PBS buffer (wherein V lipofectamine :M p-DNA =3 μL: 1 μg), stand at room temperature for 10 minutes, then add the p-DNA dispersion solution dropwise to the lipofectamine dispersion solution, and leave it at room temperature for 5 minutes. Soak the substrate in the complex solution, 500 μL solution / well, and place at 4°C for 12h.
[0024] 2) After the adsorption process was completed, the substrate was gently rinsed with PBS for 3 times and then with ultrapure water for 3 time...
Embodiment 2
[0028] 1) Diffusion depth of boron atoms is 200nm, diffusion concentration is 1×10 18 atom / cm 2 The p+ / n-type silicon wafers were cut into square pieces of 1cm×1cm and sterilized by high temperature and high pressure for later use. The process of preparing the p-DNA / lipofectamine complex is as follows: take an appropriate amount of lipofectamine and disperse it in PBS buffer, and get an appropriate amount of p-DNA and disperse it in PBS buffer (wherein V lipofectamine :M p-DNA =3 μL: 1 μg), stand at room temperature for 10 minutes, then add the p-DNA dispersion solution dropwise to the lipofectamine dispersion solution, and leave it at room temperature for 5 minutes. Soak the substrate in the complex solution, 500 μL solution / well, and place at 4°C for 12h.
[0029] 2) After the adsorption is completed, gently rinse the substrate 3 times with PBS and then gently rinse 3 times with ultrapure water, then place it in a new well plate, and inoculate cells with a cell inoculatio...
Embodiment 3
[0031] 1) Diffusion depth of boron atoms is 200nm, diffusion concentration is 1×10 18 atom / cm 2 The p+ / n-type silicon wafers were cut into 1cm×1cm square pieces. Soak the substrate in a chitosan solution (concentration: 5 mg / ml), soak for 6 hours, rinse gently with PBS for 3 times and then with ultrapure water for 3 times, sterilize with ultraviolet light for 30 minutes, and set aside.
[0032] 2) The process of preparing the p-DNA / lipofectamine complex is as follows: take an appropriate amount of lipofectamine and disperse it in PBS buffer, and take an appropriate amount of p-DNA and disperse it in PBS buffer (wherein V lipofectamine :M p-DNA =3 μL: 1 μg), stand at room temperature for 10 minutes, then add the p-DNA dispersion solution dropwise to the lipofectamine dispersion solution, and leave it at room temperature for 5 minutes. Soak the polymer layer-modified substrate prepared in step 1 in the complex solution, 500 μL solution / well, and place at 4° C. for 12 h.
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