Validation of reference genes for quantitative real-time PCR in Bupleurum chinese / 中草药

ABSTRACT

Objective To identify stable reference genes in different growth stages and different organs of Bupleurum chinese Methods All Ct values of 18 candidate internal reference genes were obtained by real-time quantitative PCR. Three software (Bestkeeper, NormFinder, and GeNorm) based on different algorithms were used to analyze the stability of the internal reference gene. The Pearson correlation coefficient was also analyzed. Results The Ct values of all candidate genes were relatively broad. ADF1b, ADF5, ADF7, eIF2b, and ACT2 were the most stable reference genes, whereas the gene of eIF6 was the least stable of the reference gene. The results of three softwares showed significant correlation. Conclusion Real-time fluorescence quantitative PCR combined with three different algorithms for the screening and validation of the reference gene of B. chinese is feasible. The homogenization of the target gene by the reference genes of the present study is helpful to improve the accuracy and reliability of gene expression analysis in molecular genetic research of B. chinese.