SEAP Expression in HeLa (Cervical Adenocarcinoma) cells with FuGENE®HD and competitors

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HeLa_HD,_6_Transit-2020,_L2k_trademarks

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.