comparative analysis of gene expression level by quantitative real-time pcr has limited application in objects with different morphology基因表达水平的比较分析定量实时聚合酶链反应限制了应用程序的对象有不同的形态.pdfVIP

Comparative Analysis of Gene Expression Level by Quantitative Real-Time PCR Has Limited Application in Objects with Different Morphology Natalia V. Demidenko1,2*, Aleksey A. Penin1,2,3 1 Department of Genetics, Biological faculty, Mikhail Vasilyevich (MV) Lomonosov Moscow State University, Moscow, Russia, 2 Evolutionary Genomics Laboratory, Faculty of Bioengineering and Bioinformatics, Mikhail Vasilyevich (MV) Lomonosov Moscow State University, Moscow, Russia, 3 A.A. Kharkevich Institute for Information Transmission Problems, Russian Academy of Science, Moscow, Russia Abstract qRT-PCR is a generally acknowledged method for gene expression analysis due to its precision and reproducibility. However, it is well known that the accuracy of qRT-PCR data varies greatly depending on the experimental design and data analysis. Recently, a set of guidelines has been proposed that aims to improve the reliability of qRT-PCR. However, there are additional factors that have not been taken into consideration in these guidelines that can seriously affect the data obtained using this method. In this study, we report the influence that object morphology can have on qRT-PCR data. We have used a number of Arabidopsis thaliana mutants with altered floral morphology as models for this study. These mutants have been well characterised (including in terms of gene expression levels and patterns) by other techniques. This allows us to compare the results from the qRT-PCR with the results inferred from other methods. We demonstrate that the comparison of gene expression levels in objects that differ greatly in their morphology can lead to erroneous results. Citation: Demidenko NV, Penin AA (2012) Comparative Analysis of Gene Expression Level by Quantitative Real-Time PCR Has Limited Application in Objects with Different Morphology. PLoS ONE 7