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Uses and Applications of FuGENE 6 Transfection Reagent

Repeat transfection, carefully pipetting FuGENE 6 Transfection Reagent directly into serum-free medium. Transfection complex was formed in medium containing serum or other additives

The FuGENE 6 / plasmid complex can be directly added to cells that are cultivated in complete medium with FCS and other additives. However, dilution of FuGENE 6 Tranfection Reagent and the formation of the complex must be done in serum-free medium without any additives. Check the original bottle of medium used for complex formation. Repeat the experiment using a new bottle of medium that does not contain any additives (e.g., serum, antibiotics, growth enhancers, etc.) Transfection complex formation time was too short

Let the FuGENE 6 Reagent/DNA complex form at least 15 minutes at room temperature. In some cell lines the transfection rate increases as the time to form the complex increases up to two hours. A suboptimal FuGENE 6 vs. DNA ratio was used Transfection efficiency is greatly reduced if the amount of DNA gets too high. Optimize the FuGENE 6 Transfection Reagent:DNA ratio according to the following procedure. Always use more FuGENE 6 Transfection Reagent (l) than DNA (g). For example, combine 3 l FuGENE 6 Transfection Reagent with 12 g DNA for a 35-mm culture dish (6-well plate).

Prepare FuGENE 6:DNA mixtures according to the following table. Do not allow FuGENE 6 Transfection Reagent to come in contact with the plastic tube before diluting with serum-free medium.

Label six tubes 1 2 3 4 5 6 Add serum-free medium (l)
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