MIQE précis: Practical implementation of minimum standard guidelines for fluorescence-based quantitative real-time PCR experiments

Fluorescence-based quantitative real-time PCR (qPCR) is commonly regarded as a straightforward, mature technology and has become a ubiquitous mainstay of molecular biology. However, obtaining, analysing and interpreting qPCR data is not a trivial issue. The progressive relegation of qPCR materials and methods to online supplements is resulting in an increasing tendency for scientific publications to contain insufficient technical information for that work to be reproduced; frequently that information is lacking altogether. Equally disconcertingly, the modest details included with publications frequently reveal the use of flawed practices that are likely to result in the publication of erroneous conclusions. This often makes it impossible for reviewers and readers to judge a manuscript's technical adequacy, whilst impeding journal editors' ability to reach publication decisions based on its validity and coherence.A set of "MIQE" guidelines proposes a minimum standard for the provision of information for publications utilising qPCR experiments [1]. These cover every aspect of sample acquisition, assay design and validation and data analysis, to provide an ideal tool for the design of de novo assays and are invaluable for guiding reviewers, as well as assisting decision-making by journal editors. However, their exhaustive incorporation of numerous assay-associated details can appear daunting to non-qPCR experts and may render them too complex for the routine reporting of established assays. Consequently, we would like to propose an abridged set of guidelines for covering key parameters of every qPCR assay (MIQE précis). These cover those steps of a qPCR assay that are essential for allowing reviewers, editors and readers to evaluate the technical merits of scientific publications utilising qPCR technology (Figure 1).As sample handling affects experimental variability, it is essential to report tissue source and to provide information about conditions of storage, partic