A detection method of 5-fluorocytosine and 5-fluorouracil in cells based on capillary electrophoresis
A technology of capillary electrophoresis and flucytosine, which is applied in the field of cell detection, can solve the problem that the separation of 5-fluorocytosine and 5-fluorouracil has not been carried out, and achieves the effects of less sample amount, short separation time and high separation efficiency.
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[0038] instrument
[0039]PA800Plus capillary electrophoresis instrument (Beckman Coulter Instruments, Fullerton, CA, USA), PDA detector, controlled by 32Krarat Station software. The effective length of the quartz capillary is 44 cm, the inner diameter of the capillary is 50 μm, and the 0.22 μm polyethersulfone membrane filter (Shanghai Anpu Experimental Technology Co., Ltd.).
[0040] 2. Cell culture
[0041] The human kidney epithelial cell line 293T was preserved and cultivated in the laboratory; DMEM dry powder, fetal calf serum, and trypsin (containing EDTA) were purchased from Gibco; the transfection reagent Lipofectamine2000 was purchased from Invitrogen; the cells were cultured in 10% serum, 1% bis Anti-(penicillin, streptomycin) DMEM medium. The cells were cultured in a 5% CO2, 37°C incubator, passaged every 2 to 3 days, and the cells in the logarithmic growth phase were used for experiments.
[0042] 3. Cell transfection
[0043] 3×10 by cell concentration 5 293...
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