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External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data
- Vermeulen, Joëlle, Pattyn, Filip, De Preter, Katleen, Vercruysse, Liesbeth, Derveaux, Stefaan, Mestdagh, Pieter, Lefever, Steve, Hellemans, Jan, Speleman, Frank, Vandesompele, Jo
- Nucleic acids research 2009 v.37 no.21 pp. e138
- computer software, gene expression, genes, nucleic acids, oligonucleotides, patients, quantitative polymerase chain reaction
- 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.