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RAW 50,000/well | ||
FuGENE® HDReagent / DNA Ratio 0.3μl/0.1μg 5μl Complex per well Complex Incubation 10 min |
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Lipofectamine® LTXReagent / DNA Ratio 0.3μl/0.1μg 20μl Complex per well Complex Incubation 30 min |
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Lipofectamine® 2000Reagent / DNA Ratio 0.5μl/0.2μg 50μl Complex per well Complex Incubation 20 min |
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Transfection with competitor reagent was performed according to manufacturer’s instructions |
RAW cells were seeded from 90-100% confluent culture the day before the transfection with the density 50,000 cells/well in 100μl complete growth medium DMEM + 10% Fetal Bovine Serum.
Tissue culture 96-round bottom well plates were used for complex preparation:
Optimal ratio reagent/DNA may vary in the range 0.25μl/0.1μg to 0.35μl/0.1μg
Transfected cells were incubated in CO2 incubator for 24 hours.