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Nucleic Acids Research Advance Access published online on September 4, 2009
Nucleic Acids Research, doi:10.1093/nar/gkp721
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© The Author(s) 2009. Published by Oxford University Press.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data

Joëlle Vermeulen1, Filip Pattyn1, Katleen De Preter1, Liesbeth Vercruysse1, Stefaan Derveaux1, Pieter Mestdagh1, Steve Lefever1, Jan Hellemans1,2, Frank Speleman1 and Jo Vandesompele1,2,*

1Center for Medical Genetics, Ghent University Hospital and 2Biogazelle, Ghent, Belgium

*To whom correspondence should be addressed. Tel: +32 9 332 5187; Fax: +32 9 332 6549; Email: joke.vandesompele{at}ugent.be

Received June 17, 2009. Revised August 2, 2009. Accepted August 15, 2009.

The quantitative polymerase chain reaction (qPCR) is widely utilized for gene expression analysis. However, the lack of robust strategies for cross laboratory data comparison hinders the ability to collaborate or perform large multicentre studies conducted at different sites. In this study we introduced and validated a workflow that employs universally applicable, quantifiable external oligonucleotide standards to address this question. Using the proposed standards and data-analysis procedure, we obtained a perfect concordance between expression values from eight different genes in 366 patient samples measured on three different qPCR instruments and matching software, reagents, plates and seals, demonstrating the power of this strategy to detect and correct inter-run variation and to enable exchange of data between different laboratories, even when not using the same qPCR platform.


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