Water-soluble fullerene nanomaterial and its preparation method and application
A nanomaterial, fullerene technology, applied in the field of biomedicine, can solve the problems of destroying the π-π conjugated system, complex chemical synthesis process, easy introduction of organic reagents, etc., to achieve improved solubility, good physical and chemical stability , save the effect of complicated process
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Embodiment 1
[0029] Example 1 Fullerene C 60 - Preparation of polydopamine-polyethyleneimine complex
[0030] 1) Take 100mg C 60 Add it into 30 mL of Tris-HCl buffer solution (pH=8.5) with a concentration of 10 mmol / L, and sonicate for 20 minutes to disperse evenly. Then, 200 mg of dopamine hydrochloride was added to the above system, and stirred for 10 h at 25° C. in the dark, at a speed of 1000 r / min.
[0031] 2) The above product was centrifuged at 8000r / min for 5 minutes, washed with distilled water and absolute ethanol, respectively, until the upper centrifuge was transparent, and the lower precipitate was placed in a vacuum drying oven for 24 hours, and the temperature was set at 40°C.
[0032] 3) Take the above dried fullerene C 60 - 100 mg of polydopamine product and 200 mg of polyethyleneimine (molecular weight: 600) were added to 20 mL of sodium hydroxide solution with a concentration of 0.1 mol / L, stirred and reacted at 25°C for 10 h, and the rotation speed was 1000 r / min.
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Embodiment 2
[0037] Example 2 Fullerene C 60 - Light-induced DNA shearing effect test of polydopamine-polyethyleneimine complex.
[0038] 1) Take 2.5 μL of 50ng / μL pSP72 plasmid DNA and mix with equal volumes of fullerene C at different concentrations 60 - Aqueous solutions of polydopamine-polyethyleneimine (125, 62.5, 31.25, 15.625, 7.8125 and 3.91 μg / ml) were mixed well.
[0039] 2) Under the condition of visible light irradiation, co-incubate at room temperature for 2 hours.
[0040] 3) Add 1 uL of 6X loading buffer (40% (w / v) sucrose, 0.25% bromophenol blue and 0.25% xylene cyanol) to the above sample 2) respectively and mix well.
[0041] 4) Add the above samples respectively 3) to the sample wells of 1% (w / v) agarose gel, and wash in 1X TAE electrophoresis buffer (40mmol / L Tris, 20mmol / L acetic acid, 1mmol / L EDTA, pH 8.0), electrophoresis was performed at 100V for 30min.
[0042] 5) After electrophoresis, the gel was taken out, stained in 10 mg / mL ethidium bromide solution for 5 ...
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