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BMC Genomics

, 6:59

First Online: 27 April 2005Received: 06 October 2004Accepted: 27 April 2005

Abstract

BackgroundThe use of microarray technology to assess gene expression levels is now widespread in biology. The validation of microarray results using independent mRNA quantitation techniques remains a desirable element of any microarray experiment. To facilitate the comparison of microarray expression data between laboratories it is essential that validation methodologies be critically examined. We have assessed the correlation between expression scores obtained for 48 human genes using oligonucleotide microarrays and the expression levels for the same genes measured by quantitative real-time RT-PCR qRT-PCR.

ResultsCorrelations with qRT-PCR data were obtained using microarray data that were processed using robust multi-array analysis RMA and the MAS 5.0 algorithm. Our results indicate that when identical transcripts are targeted by the two methods, correlations between qRT-PCR and microarray data are generally strong r = 0.89. However, we observed poor correlations between qRT-PCR and RMA or MAS 5.0 normalized microarray data for 13% or 16% of genes, respectively.

ConclusionThese results highlight the complementarity of oligonucleotide microarray and qRT-PCR technologies for validation of gene expression measurements, while emphasizing the continuing requirement for caution in interpreting gene expression data.

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2164-6-59 contains supplementary material, which is available to authorized users.

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Author: Peter B Dallas - Nicholas G Gottardo - Martin J Firth - Alex H Beesley - Katrin Hoffmann - Philippa A Terry - Joseph R

Source: https://link.springer.com/article/10.1186/1471-2164-6-59



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