实时荧光定量PCR中内参基因的选择

Selection of Reference Genes for Real-time Quantitative PCR

摘要:

实时荧光定量PCR技术是分析基因表达谱的一种常用方法, 在分析中选择合适的内参基因对数据进行校正是得到可信数据的关键。以Lactobacillus helveticus H9为研究对象, 应用实时荧光定量PCR技术, 评价了5种常用内参基因ldh、recA、rpoB、gapdh和16S rRNA的表达稳定性, 通过geNorm和NormFinder程序进行数据分析, 结果表明5个候选内参基因在菌株不同的发酵时间点表达相对都较为稳定, 结合两种分析得到其中最为稳定的基因是ldh, 适合于用作后续实时荧光定量PCR试验中的内参基因。

Abstract:

Real-time quantitative PCR is a commonly used method to analyse the gene expression profile, it is important to select an appropriate reference gene for normalization of experimental data when using this method. In our study, we used two statistical methods to evaluate the gene expression stabilities of five reference genes (ldh, recA, rpoB, gapdh and 16S rRNA) under the different growth phases of Lactobacillus helveticus H9. The results showed that the best reference gene was ldh which was the most stable gene would be used for normalization of real-time quantitative PCR experiments data.