Pfaffl Method
The Pfaffl model generalises 2^-ddCt to unequal efficiencies as ratio = E_target^dCq(target) / E_reference^dCq(reference), using each assay's measured efficiency instead of assuming 2. It is the correct choice whenever standard curves show target and reference efficiencies differ meaningfully, and it reduces to 2^-ddCt when both equal 2.
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The Pfaffl model generalises 2^-ddCt to unequal efficiencies as ratio = E_target^dCq(target) / E_reference^dCq(reference), using each assay's measured efficiency instead of assuming 2. It is the correct choice whenever standard curves show target and reference efficiencies differ meaningfully, and it reduces to 2^-ddCt when both equal 2.
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