Samstag, 7. Januar 2012

A MIQE Case Study — Effect of RNA Sample Quality and Reference Gene Stability on Gene Expression Data



A MIQE Case Study — Effect of RNA Sample Quality and Reference Gene Stability on Gene Expression Data


by Sean Taylor, Bio-Rad Laboratories Canada, 1329 Meyerside Dr Mississauga, ON, L5T 1C9, and Marguerite Buchanan and Mark Basik, McGill University, Jewish General Hospital, Montreal, QC, published: December 15, 2011

Abstract - Real-time quantitative PCR (qPCR) has become the gold standard for validating DNA microarray data and is routinely used to determine gene expression differences between a wide variety of samples. The exquisite sensitivity of the technology permits the detection of a single copy of a target gene in a sample which has led to qPCR now being used in the clinical setting to diagnose infection and disease states. In an effort to standardize the design of the associated experiments, the minimum information for publication of quantitative real-time PCR experiments (MIQE) guidelines were published in 2009. In this study, we show how qPCR can lead to erroneous conclusions regarding differences in MCM7 gene expression between normal and tumor human breast cancer samples if the key steps set out in the MIQE guidelines are not followed.

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