qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data 英文参考文献.docVIP

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qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data 英文参考文献.doc

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qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data 英文参考文献

Open Access Me d 2007HellemansetVoalu.metho8, Issue 2, Article R19 qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data Jan Hellemans, Geert Mortier, Anne De Paepe, Frank Speleman and Jo Vandesompele Address: Center for Medical Genetics, Ghent University Hospital, De Pintelaan, B-9000 Ghent, Belgium. Correspondence: Jo Vandesompele. Email: Joke.Vandesompele@UGent.be Published: 9 February 2007 Received: 31 August 2006 Revised: 7 December 2006 Accepted: 9 February 2007 Genome Biology 2007, 8:R19 (doi:10.1186/gb-2007-8-2-r19) The electronic version of this article is the complete one and can be found online at /2007/8/2/R19 ? 2007 Hellemans et al.; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. AutomatedpqBase, aanalysisfree proofgrreal-timeam for theqPCRmanadatagement and automated analysis of qPCR data, is described/p Abstract Although quantitative PCR (qPCR) is becoming the method of choice for expression profiling of selected genes, accurate and straightforward processing of the raw measurements remains a major hurdle. Here we outline advanced and universally applicable models for relative quantification and inter-run calibration with proper error propagation along the entire calculation track. These models and algorithms are implemented in qBase, a free program for the management and automated analysis of qPCR data. Background number of replicates, normalization using only one reference gene, lack of data quality controls (for example, replicate var- iability, negative controls, reference gene expression stabil- ity), inability to calibrate multiple runs, limited result visualization options, lack