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We always suggest our customers to replace cell media by adding fresh media without antibiotic (pen/strep) at the time of transfecting cells.
We also have done some studies with and without antibiotic-containing cell media during regular DNA transfection procedures. We have found that antibiotics (pep/strep) do not affect the ability of lipofectamine reagents to transfect cells. However, we have not done a comparison for Lentiviral production.
Infection the target cells with lentivirus, the cell culture media can be in presence or absence pep/strep antibiotic.