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Yes, antibiotics can be used during transfection with any of the Life Technologies lipid based reagents. However, the critical steps of the protocol need to be performed with the recommended OptiMEM Media and not culture media that contains any serum or antibiotics. The transfection protocol involves a two tube protocol and It is important to add the lipid reagent to OptiMEM in one tube and the DNA or RNA to OptiMEM in another tube and mix each tube thoroughly. Then, equal amounts from each tube should be mixed to form the transfection complex and incubated for five minutes at room temperature. The lipid-DNA or lipid-RNA complex can then be added directly to cells in complete culture media. Analysis of transfection results can be performed 24-48 hours post transfection.