Selection of suitable reference genes for real-time PCR studies of early developmental stages of sturgeons

In quantitative real-time PCR¡ the mRNA level can be quantified in relative terms based on the expression ratio of mRNAs of the target gene and an internal reference gene. Since¡ an internal standard should be expressed at a constant level among different tissues of an organism at all stages of development¡ and should be unaffected by the experimental treatment¡ the stability of different reference genes needs to be tested for every new experiment to select the most suitable reference gene for normalization of target gene. Here in this study¡ the stability of six candidate reference genes including ribosomal protein L6 (RPL6)¡ ribosomal protein L13 (RPL13)¡ Elongation factor 1-alpha (EF1A)¡ Ubiquitin (UBQ)¡ Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and beta-actin (ACTB) were studied during early development of Persian sturgeon¡ Acipenser persicus using Real-Time PCR TaqMan. The general consensus from BestKeeper¡ NormFinder and geNorm models indicate that RPL6 and ACTB were found to be the most stable reference genes across the different developmental time-points¡ whereas RPL13 was the most variable gene. Using RPL6¡ ACTB and a two-gene normalization factor based on the geometric average of RPL6/ACTB could be recommended for standardization of qPCR data in future developmental studies of Persian sturgeon and likely in other Acipenserid species.