Brief Funct Genomics. 2018 May 1;17(3):147-150. doi: 10.1093/bfgp/elx022.
The performance of the quantitative polymerase chain reaction (qPCR) assay in the analysis of gene expression belonging to multigene families in tissues rich in secondary metabolites is technically complicated. Here, we present the qPCR analysis of PMT2 gene, a predominant member of a multigene family from tobacco, expressed in the root tissues and is involved in the biosynthesis of nicotine. Consequently, we provide insight into the effect of polymerase chain reaction (PCR) amplification efficiency (AE) of reference and target genes of calibrator and test samples on the qPCR assay performance. Obviously, we found PCR AE as a critical indicator of qPCR assay performance involving multigene families and secondary metabolite-rich root tissues of tobacco. The integration of consistent and uniform PCR amplification efficiencies of reference and target genes of the samples into the relative quantification analysis is emphasized.
定量聚合酶链反应（qPCR）分析在富含次生代谢产物的组织中分析多基因家族的基因表达时，技术上较为复杂。在这里，我们介绍了 qPCR 分析 PMT2 基因，该基因为烟草多基因家族的主要成员，在根组织中表达，并参与尼古丁的生物合成。因此，我们深入了解了参比和测试样品的聚合酶链反应（PCR）扩增效率（AE）对 qPCR 分析性能的影响。显然，我们发现 PCR AE 是涉及多基因家族和富含次生代谢产物的烟草根组织的 qPCR 分析性能的关键指标。我们强调将参比和测试样品的目标基因的一致和均匀的 PCR 扩增效率整合到相对定量分析中。
