12097期排列5 www.ixpcv.com Sinofection? is a superior cationic polymer-based transfection reagent. Compared with liposomes on the market, Sinofection exhibits lower cytotoxicity, higher transfection efficiency and higher repeatability.
|The transfection procedures of adherent cells are shown in Fig. 1.
we take the 96-well plates as example：
4cells/well (table 1)and cultured with 5%CO2 at 37℃ for 18-24h before transfection.
2. Preparation of transfection mixture:
(1). Dilute DNA into serum-free medium(25μL in total) and homogenize gently.
(2). Dilute Sinofection into serum-free medium(25μL in total) and homogenize gently and incubate at room temperature for 5 min.
(3). Mix the DNA and Sinofection at room temperature for 15~20min 3. Remove the culture medium and add 50μL mixture per well.
4. After 4-6hours(for SF9 cell is 2h), remove the transfection mixture and add medium with serum.
5. Gene expression is tested after incubation with 5% CO2 at 37℃ for 48-72h(incubate sf9 cell line at 27℃ for 48-72h).
Fig. 1 Adherent cells' Transfection
(1). The experiment conditions for different cell lines are shown in Table 1. The corresponding transfection protocols can be got by entering links in Table 1.
(2). Number of cells per well, dosage of Sinofection, DNA and serum-free medium for dilution are proportional to basal area per well for plates of different sizes. The basical areas for plates with different sizes are shown in Table 2. The dosage and ratio of DNA and Sinofection should be optimized to achieve the best transfection results.
Table1. Usage of Sinofection in different cell lines（96-well plate）
|Cell type||Culture medium||Cells per well||DNA||Sinofection||Medium change after 4-6h|
|MCF7||MEM/NEAA+0.01mg/mL insulin + sodium pyruvat||2×104||0.1μg||0.25μL||MEM/NEAA+0.01mg/mL insulin + sodium pyruvat+10%FBS|
|sf9||SIM SF||5×104||0.4μg||0.75μL||SIM SF+10%FBS|
Table 2. Scaling up or down transfections with Sinofection（according to the growth area）.
Note: the actual conditions should be optimized by experiments.