Cargando…
External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data
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 introd...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2009
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2790878/ https://www.ncbi.nlm.nih.gov/pubmed/19734345 http://dx.doi.org/10.1093/nar/gkp721 |
_version_ | 1782175143095697408 |
---|---|
author | Vermeulen, Joëlle Pattyn, Filip De Preter, Katleen Vercruysse, Liesbeth Derveaux, Stefaan Mestdagh, Pieter Lefever, Steve Hellemans, Jan Speleman, Frank Vandesompele, Jo |
author_facet | Vermeulen, Joëlle Pattyn, Filip De Preter, Katleen Vercruysse, Liesbeth Derveaux, Stefaan Mestdagh, Pieter Lefever, Steve Hellemans, Jan Speleman, Frank Vandesompele, Jo |
author_sort | Vermeulen, Joëlle |
collection | PubMed |
description | 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. |
format | Text |
id | pubmed-2790878 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2009 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-27908782009-12-09 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 Res Methods Online 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. Oxford University Press 2009-11 2009-09-04 /pmc/articles/PMC2790878/ /pubmed/19734345 http://dx.doi.org/10.1093/nar/gkp721 Text en © The Author(s) 2009. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/2.5/uk/ 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. |
spellingShingle | Methods Online Vermeulen, Joëlle Pattyn, Filip De Preter, Katleen Vercruysse, Liesbeth Derveaux, Stefaan Mestdagh, Pieter Lefever, Steve Hellemans, Jan Speleman, Frank Vandesompele, Jo External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data |
title | External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data |
title_full | External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data |
title_fullStr | External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data |
title_full_unstemmed | External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data |
title_short | External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data |
title_sort | external oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative pcr data |
topic | Methods Online |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2790878/ https://www.ncbi.nlm.nih.gov/pubmed/19734345 http://dx.doi.org/10.1093/nar/gkp721 |
work_keys_str_mv | AT vermeulenjoelle externaloligonucleotidestandardsenablecrosslaboratorycomparisonandexchangeofrealtimequantitativepcrdata AT pattynfilip externaloligonucleotidestandardsenablecrosslaboratorycomparisonandexchangeofrealtimequantitativepcrdata AT depreterkatleen externaloligonucleotidestandardsenablecrosslaboratorycomparisonandexchangeofrealtimequantitativepcrdata AT vercruysseliesbeth externaloligonucleotidestandardsenablecrosslaboratorycomparisonandexchangeofrealtimequantitativepcrdata AT derveauxstefaan externaloligonucleotidestandardsenablecrosslaboratorycomparisonandexchangeofrealtimequantitativepcrdata AT mestdaghpieter externaloligonucleotidestandardsenablecrosslaboratorycomparisonandexchangeofrealtimequantitativepcrdata AT lefeversteve externaloligonucleotidestandardsenablecrosslaboratorycomparisonandexchangeofrealtimequantitativepcrdata AT hellemansjan externaloligonucleotidestandardsenablecrosslaboratorycomparisonandexchangeofrealtimequantitativepcrdata AT spelemanfrank externaloligonucleotidestandardsenablecrosslaboratorycomparisonandexchangeofrealtimequantitativepcrdata AT vandesompelejo externaloligonucleotidestandardsenablecrosslaboratorycomparisonandexchangeofrealtimequantitativepcrdata |