GFP expression in T98G (Human Brain Glioblastoma) cells, 48 hours after transfection by FuGENE® HD and Competitor Lipofectamine® 2000, 15,000 cells/well

Complex formed in OptiMEM®.

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Transfection is done in
100% Fetal Bovine Serum
Trasfection is done in complete growth MEM medium supplemented with 1 mM Sodium Piruvate , 10 mM MEM non-essential Amino Acids Solution and 10% Fetal Bovine Serum
Reagent / DNA Ratio
0.4μl/0.1μg
8μl Complex per well
Complex Incubation 5 min

FuGENE®HD

 

Reagent / DNA Ratio
0.4μl/0.1μg
8μl Complex per well
Complex Incubation 5 min
Reagent / DNA Ratio
0.5μl/0.2μg
50μl Complex per well
Complex Incubation 20 min

 Lipofectamine® 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 T98G (Human Brain Glioblastoma) Cell Transfection by FuGENE HD with gWIZ™ GFP-plasmid.

  1. Cell plating

    T98G cells were seeded from 90-100% confluent culture the day before transfection with the density 15,000 cells/well in 100 μl complete growth MEM medium supplemented with 1 mM Sodium Piruvate, 10 mM MEM non-essential Amino Acids Solution and 10% Fetal Bovine Serum.

    For transfection in 100% Fetal Bovine Serum the complete growth medium was replaced with Fetal Bovine Serum two hours before transfection.

  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®.
    2. Add 8 μl of reagent to 160 μl of OptiMEM® / 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.
  3. Incubation

    Place the cells into CO2 incubator for 48 hours.

  4. Detection of GFP expression

    Monitor the transgene expression with fluorescent microscope or flowcytometer.