Screening of Reference Genes for Gene Expression in Layer Hens Tissues Using Real-time Quantitative PCR

To screen the reference genes of stable expression in different tissues of Hy-line layer (Gallus domesticus), RPS2, β-actin, GAPDH and HMBS genes were chosen as candidate gene, the software of geNorm, NormFinder and the Ct value were employed to analyze their stability in eight tissues (heart, liver, lung, kidney, duodenum, tibia, pancreas and uterus) of 120 days of age layers using Real-time quantitative PCR technology. The results showed that RPS2 gene was always dominant stability by the analysis of Ct value and geNorm software, the β-actin gene was followed; The NormFinder software showed that the HMBS gene was relative stability, and HMBS and RPS2 genes were the optimal combination; When analyzed the expression of constant reference gene in different tissues, it found that the most stable reference gene in different tissues was not the same. Therefore, it concluded that different tissues had different optimal gene, but they had the common potential, RPS2 and β-actin genes had the relative expression stability comparing the other gene.