Evaluation of reliable reference genes for the lung of neonatal C57BL/6 mice under consideration of biological, technical and experimental variables

A pool of reference gene candidates was first gathered by reviewing previous studies and a commercial panel as well as using Genevestigator, analysis software for published microarray experiments. In the next step, the expression of the potential reference gene candidates was evaluated by the present microarray from the neonatal C57BL/6 mice with otherwise physiologic conditions and through this process, the 24 final reference gene candidates were chosen. Then, RT-qPCR validation of the 24 reference gene candidates and data analysis by GeNorm and Normfinder were completed, so that reliable reference genes specific for biological, technical and experimental conditions were suggested. Microarray experiment was performed in the lung tissue of three neonatal (5-7-day-old) C57BL/6 mice from physiologic conditions (FiO2=0.21). Each mouse was investigated in two technical replicates.