AKT1 and breast neoplasm: Since the assay had not been validated using FFPE samples, we first set out to establish the threshold values for the mutation assay Ct, reference assay Ct, and ΔCt for AKT1 E17K mutation detection in FFPE samples using the AKT1 E17K FFPE DNA reference standards and included DNA from 10 AKT1 wild-type FFPE breast tumors as a negative control.