In order for infection to be most effective, you must optimize transfection efficiencies at this step to preserve library complexity. Upon infecting the Tet cell line with the virus, you create a pool of cells containing the high-complexity cDNA library. After infection, add or remove Dox to activate transcription of the library insert. To illustrate this central feature, we infected HeLa Tet-Off Cells with the library. After infection, you can subject your cells to any appropriate screen or selection scheme to isolate positive clones and create a stable cell line expressing the desired insert.
Positive cells stain brown.
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