HeLa cells morphology 24 hours after transfection by FuGENE®HD, FuGENE®6 and competitor Lipofectamine® 2000

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Untransfected Cells Reagent / DNA Ratio
10μl Complex per well
Complex Incubation 15 min


Lipofectamine® 2000

Reagent / DNA Ratio
8μl Complex per well
Complex Incubation 5 min

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

Protocol for HeLa Cell Transfection by FuGENE HD

  1. Cell plating

    HeLa cells were seeded from 90-100% confluent culture the day before transfection with the density 10,000 cells per well in 100 μl complete growth medium DMEM + 10% Fetal Bovine Serum.

  2. Complex preparation (per 20 wells)

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

    1. Prepare pCMVβ DNA solution in OptiMEM® or sterile deionized water, 0.0125μg/μl.
    2. Add 6 μl of reagent to 160 μl 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 ratio Reagent/DNA may vary in range 0.25μl/0.1μg to 0.35μl/0.1μg.

  3. Incubation

    Place the cells into CO2 incubator for 24 hours.