GFP expression in HEK 293 cells transfected with FuGENE®HD, 72 hours incubation, 50,000 cells/well plated.

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Complexes are formed in OptiMEM®

FuGENE®HD

FuGENE HD
Reagent / DNA Ratio
0.4μl/0.1μg
8μl Complex per well
Complex Incubation 5 min
 

*An important carrier line for virus production

Protocol for HEK293 Cell Transfection by FuGENE HD

  1. Cell plating

    Cells were seeded in 96-well non-tissue culture plate just before transfection with the density 50,000 cells per well in 100 μl “FreeStyle™ 293” growth medium (Invitrogen, Inc.®).

  2. Complex preparation (per 20 wells)

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

    1. Prepare 0.0125μg/μl gWIZ™ GFP DNA solution in OptiMEM® or sterile deionized water.
    2. Add 8 μl of FuGENE®HD to 160μl of DNA solution.
    3. Mix carefully by pipetting (15 times).
    4. Incubate 5 min at room temperature.
    5. Add 8 μl of complex per well to the cells, and mix thoroughly.

    Optimal reagent/DNA ratio may vary in the range 0.25μl/0.1μg to 0.45μl/0.1μg.

  3. Incubation

    Place the cells into CO2 incubator for 72 hours.

  4. Detection of GFP expression

    Monitor the transgene expression under fluorescent microscope or flowcytometer.