Attention: 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 Clarkfection, 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 Clarkfection should be optimized to achieve the best transfection results. Table1. Usage of Clarkfection in different cell lines(96-well plate) Cell type | Culture medium | Cells per well | DNA | Clarkfection | Medium change after 4-6h | 293H | DMEM | 3×104 | 0.2µg | 0.5µL | DMEM+10%FBS | 293FT | DMEM | 3×104 | 0.2µg | 0.5µL | DMEM+10%FBS | 293E | DMEM | 3×104 | 0.2µg | 0.5µL | DMEM+10%FBS | 293F | DMEM | 3×104 | 0.2µg | 0.5µL | DMEM+10%FBS | COS7 | DMEM | 1.5×104 | 0.4µg | 0.5µL | DMEM+10%FBS | hela | DMEM | 2×104 | 0.3µg | 0.5µL | 1640+15%FBS | Caco2 | MEM | 3.5×104 | 0.3µg | 0.75µL | MEM+10%FBS | BHK21 | MEM | 2×104 | 0.2µg | 0.5µL | MEM+10%FBS | CHO-DG44 | DMEM+HT+pro | 2×104 | 0.5µg | 0.5µL | DMEM+HT+pro +10%FBS | RAW264.7 | DMEM | 3×104 | 0.2µg | 0.5µL | DMEM +10%FBS | 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 | SW480 | IMDM | 3×104 | 0.4µg | 0.5µL | IMDM +10%FBS | MDCK | DMEM | 4×104 | 0.6µg | 1µL | DMEM+10%FBS | CHO-K1 | IMDM+Pro | 3×104 | 0.2µg | 0.5µL | IMDM+Pro +10%FBS | HepG2 | DMEM | 3×104 | 0.5µg | 0.75µL | DMEM+10%FBS | A549 | DMEM | 2×104 | 0.3µg | 0.5µL | DMEM+10%FBS | NIH/3T3 | DMEM | 1.5×104 | 0.1µg | 0.75µL | DMEM+10%FBS | vero | DMEM | 3×104 | 0.3µg | 0.75µL | DMEM+10%FBS | sf9 | SIM SF | 5×104 | 0.4µg | 0.75µL | SIM SF+10%FBS | Table 2. Scaling up or down transfections with Clarkfection(according to the growth area). Note: the actual conditions should be optimized by experiments.  |