GFP expression in SNU-16 (ATCC®: CRL-5974™ Human Gastric Carcinoma) cells 72 hours after transfection by FuGENE®HD and Reagents from Competitor in 96 well plates, 10,000cells/well

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Transfection is done in RPMI 1640+10% Fetal Bovine Serum  Cell Morphology

Reagent / DNA Ratio
0.4μl/0.1μg
5μl Complex per well
Complex Incubation 10 min
Complexes are formed in OptiMEM®

FuGENE®HD

 


Reagent / DNA Ratio
0.4μl/0.1μg
5μl Complex per well
Complex Incubation 10 min

Reagent / DNA Ratio
0.4μl/0.1μg
5μl Complex per well
Complex Incubation 10 min
Complexes are formed in Sterile Deionized Water

FuGENE®HD

 


Reagent / DNA Ratio
0.4μl/0.1μg
5μl Complex per well
Complex Incubation 10 min

Reagent / DNA Ratio
0.5μl/0.2μg
50μl Complex per well
Complex Incubation 20 min
Complexes are formed in OptiMEM®

Lipofectamine® 2000

 


Reagent / DNA Ratio
0.5μl/0.2μg
50μl Complex per well
Complex Incubation 20 min

Reagent / DNA Ratio
0.3μl/0.1μg
20μl Complex per well
Complex Incubation 30 min
Complexes are formed in OptiMEM®

Lipofectamine® LTX

 


Reagent / DNA Ratio
0.3μl/0.1μg
20μl Complex per well
Complex Incubation 30 min
Transfections with competitor reagents were done according to manufacturer’s instructions.

Protocol for transfection of SNU-16 (ATCC®: CRL5974™) Cells by FuGENE HD in 96 well plates with gWIZ™ GFP plasmid.

  1. Cell plating

    SNU-16 cells were seeded from 90-100% confluent culture the day before transfection with the density 10,000 cells/well in 100 μl complete growth medium (RPMI + 10% Fetal Bovine Serum).

  2. Complex preparation (per 20 wells)

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

    1. Prepare 0.02μg/μl gWIZ™ GFP DNA solution in OptiMEM® or sterile deionized water.
    2. Add 8 μl of reagent to 100 μl of DNA solution.
    3. Mix carefully by pipetting (15 times).
    4. Incubate 10 min at room temperature.
    5. Add 5 μl of complex per well to the cells, and mix thoroughly.
  3. Incubation

    Transfected cells were incubated in CO2 incubator for 72 hours.

  4. Detection of GFP expression

    Monitor the transgene expression under fluorescent microscope or flowcytometer.