Recently I have tried to transfect mRNA (8kb) into cells using lipofectamine 2000 in serum free media. However, the transfection efficiency is very low. Why do some transfection protocols use serum containing media? (Note: the mRNA-lipofectamine complex formed in serum free opti-MEM).
Dear Tan Chee,
Did you find any solution for transfecting mRNA into cells ?
Transfection protocols recommend not using serum or supplement because these supplements (proteins, antibiotics, growth factors...) can interfere with the formation of complexes. Nucleic acids (DNA, siRNA, mRNA...) will be in competition with all supplment to form lipoplexes. Moreover, there is a high risk of degradation in complete serum if nucleic acids are not well protected.
Once complexes are formed (of course depending on the transfection reagent), you can add them into complete medium without any loss of efficiency.
Hope it will help;
best,
Cedric
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