Selection of reference genes for real-time fluorescence quantitative PCR in Nerviliao Fordii Folium / 中草药

ABSTRACT

Objective: To select an appropriate reference gene for gene expression analysis using real-time fluorescence quantitative PCR (qRT-PCR). Methods: Using leaf, petiole, and corm tissues of N. fordii, five common reference genes (18S rRNA, actin, ubiquitin, EF-1α, and β-tubulin) were compared. The stability of the candidate reference genes was evaluated by Ct value using GeNorm and NormFinder software. Results: The stabilities of five reference genes varied from each other in three tissues of N. fordii. The analysis of GeNorm and NormFinder exhibited that the stability of β-tubulin was the most stable, followed by EF-1a, Ubiquitin, actin, and 18S rRNA in order. Conclusion: The β-tubulin gene could be served as the reference gene for the normalization of gene expression in different tissues of N. fordii using qRT-PCR.