, Ct sample assay (x) -Ct sample assay (ext ctrl) = dCt sample assay (x) 2. Normalize using median IPC (per experiment): dCt sample assay (x) -Median dCt IPC assay (x) =

, Calculate average value for background (per experiment): Average (ddCt background assay(x) ) = Bg IPC (x)

, Calculate median value for the background of all assays: Median (Bg IPC (x) )