CRA-Centro di Ricerca per l'Enologia, Via Pietro Micca 35, Asti, Italy.
Biotechnol Lett. 2011 Aug;33(8):1593-9. doi: 10.1007/s10529-011-0603-y. Epub 2011 Mar 31.
Expression data from RT-qPCR (reverse transcription quantitative PCR) needs to be normalized to account for experimental variability among samples caused by differential yields of the transcripts in RNA extraction or in the reverse transcription. The most common method is to normalize against one or more reference genes (RG). We have selected RGs suitable for normalization of RT-qPCR raw data in Saccharomyces cerevisiae during alcoholic fermentation. The RGs were evaluated by three different statistical methods. The suitability of the selected RG sets was compared with ACT1, a commonly used non-validated single RG, by normalizing the expression of two target genes. Expression profiles of the target genes revealed the risk of misleading interpretation of expression data due to an unreliable RG.
从 RT-qPCR（逆转录定量 PCR）获得的表达数据需要进行标准化，以考虑到 RNA 提取或逆转录过程中转录本产量的差异引起的样品间实验变异性。最常用的方法是针对一个或多个参考基因（RG）进行归一化。我们已经选择了适合酿酒酵母酒精发酵过程中 RT-qPCR 原始数据归一化的 RG。通过对两个靶基因的表达进行归一化，使用三种不同的统计方法评估 RG 的适用性。通过比较选定 RG 集与常用的未经验证的单个 RG ACT1 的归一化，评估了选定 RG 集的适用性。靶基因的表达谱揭示了由于 RG 不可靠而导致对表达数据的解释产生误导的风险。
