%0 Generic %A X., Cui %A J., Zhou %A J., Qiu %A M.R., Johnson %A M., Mrug %D 2009 %T Supplementary Material for: Validation of Endogenous Internal Real-Time PCR Controls in Renal Tissues %U https://karger.figshare.com/articles/dataset/Supplementary_Material_for_Validation_of_Endogenous_Internal_Real-Time_PCR_Controls_in_Renal_Tissues/5120821 %R 10.6084/m9.figshare.5120821.v1 %2 https://karger.figshare.com/ndownloader/files/8705143 %K Real-time PCR %K Kidney disease %K Endogenous internal control %K Reference gene %K Housekeeping gene %X Background: Endogenous internal controls (‘reference’ or ‘housekeeping’ genes) are widely used in real-time PCR (RT-PCR) analyses. Their use relies on the premise of consistently stable expression across studied experimental conditions. Unfortunately, none of these controls fulfills this premise across a wide range of experimental conditions; consequently, none of them can be recommended for universal use. Methods: To determine which endogenous RT-PCR controls are suitable for analyses of renal tissues altered by kidney disease, we studied the expression of 16 commonly used ‘reference genes’ in 7 mildly and 7 severely affected whole kidney tissues from a well-characterized cystic kidney disease model. Expression levels of these 16 genes, determined by TaqMan® RT-PCR analyses and Affymetrix GeneChip® arrays, were normalized and tested for overall variance and equivalence of the means. Results: Both statistical approaches and both TaqMan- and GeneChip-based methods converged on 3 out of the 4 top-ranked genes (Ppia, Gapdh and Pgk1) that had the most constant expression levels across the studied phenotypes. Conclusion: A combination of the top-ranked genes will provide a suitable endogenous internal control for similar studies of kidney tissues across a wide range of disease severity. %I Karger Publishers