GFP expression in CHO-S cells transfected by FuGENE®HD and Competitor Lipofectamine® 2000, 72 hours incubation, 50,000 cells/well plated

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Complexes are formed in OptiMEM®

FuGENE®HD

FuGENEHD
Reagent / DNA Ratio
0.45μl/0.1μg
8μl Complex per well
Complex Incubation 0 min

Lipofectamine® 2000

LFA 2000
Reagent / DNA Ratio
0.5μl/0.2μg
50μl Complex per well
Complex Incubation 20 min
 Transfection with competitor reagent was performed according to manufacturer’s instructions

Protocol for CHO-S Cell Transfection by FuGENE HD

  1. Cell plating

    Cells for transfection were grown in 15 mL CD-CHO medium (Gibco®) in 50 ml tubes on rotator at 110 RPM. The rotor was placed at 40° angle. Cells for transfection were harvested at approximately 1,000,000 cells / ml. Cells were seeded in 96-well flat bottom non-tissue culture plate just before transfection with the density 50,000 cells per well in 100 μl growth medium.

  2. Complex preparation (per 20 wells)

    Tissue culture 96-round bottom well plates were used for complex preparation:

    1. Prepare 0.0125 μg/μl gWIZ™ GFP DNA solution in sterile deionized water.
    2. Add 9 μl of FuGENE®HD to 160μl of DNA solution.
    3. Mix carefully by pipetting (10-15 times).
    4. Incubation of complex is not necessary.
    5. Add 8 μl of complex to the cells, per well and mix thoroughly.

    Optimal FuGENE®HD/DNA ratio may vary in range 0.3μl/0.1μg to 0.5μl/0.1μg.

  3. Incubation

    Place the cells into CO2 incubator with orbital shaker at 110 RPM for 72 hours.

  4. Detection of GFP expression

    Monitor the GFP expression under fluorescent microscope or by flow cytometery.