Screening of the reference genes of Skeletonema marinoi under different concentration of Fe 3+ conditions in real-time quantitative PCR analysis

Abstract

Real-time quantitative PCR (qRT-PCR) is a common method for quantitative analysis of gene expression. Selection of appropriate reference genes is essential for the accurate analysis of target gene expression levels. In this study, the expression of seven reference genes of Cytb, EF-1α, HPRT, UBC, GAPDH, β-actin and β-tubulin was quantitatively analyzed with different concentrations of iron concentration. The GeNorm, NormFinder and BestKeeper software comprehensively evaluated the stability of these reference genes. The results showed that the expression stability of Cytb and EF-1α was better, and the combination of EF-1α + Cytb was the best. It could be used as a reference gene for the study of gene expression in Skeletonema marinoi, while the expression stability of other genes was poor, and they were not suitable for being used as a reference gene. This study provides a methodological basis for the selection of reference genes during the study of gene expression in S. marinoi.