%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