Can we trust your published qPCR data?
The pressure to publish is causing shortcuts which result in misleading papers being published.
Today there is a great deal of pressure to publish papers in high-ranking, high-impact factor journals in order to make it possible to write successful grant applications. The initial purpose of publishing is overshadowed by the pressure of having as many papers as possible in high-impact factor publications. This has a significant impact on the integrity of the generated data which is too often non-reproducible. Obviously, a paper should enable anyone with the right competencies to repeat what has been done, and the main aim of the paper should be to transmit this information in an understandable way.
We have the means to get lots of data using dPCR, Microarrays, NGS, qPCR. The data needs to be published so the hypotheses are being established around the data. Instead of being hypothesis driven, the research now tends to be data driven. The hypotheses are established to fit the data.
Professor Stephen A. Bustin, Faculty of Medical Science, Anglia Ruskin University, UK
WHY MIQE – The Minimum Information for Publication of Quantitative Real-Time PCR Experiments?
Because there is a lack of a consensus on how to perform and interpret qPCR experiments and because there is insufficient experimental detail in many publications. The MIQE guidelines target the reliability of results to help ensure the integrity of the scientific literature, promote consistency between laboratories, and increase experimental transparency. By providing all relevant experimental conditions and assay characteristics, reviewers can assess the validity of the protocols used. Full disclosure of all reagents, sequences, and analysis methods is necessary to enable other investigators to reproduce results.
”In terms of compliance with 16 critical parameters that allow any competent reader to reproduce data, the information is simply not provided in the vast majority of papers using qPCR. ” Professor Stephen A. Bustin, Faculty of Medical Science, Anglia Ruskin University, UK
MIQE & qPCR iBook
The iBook offers detailed insight into experimental design, sampling, RNA quality control, reverse transcription, qPCR instrumentation, qPCR assay validation, quality control, quantification strategies, reference genes selection, qPCR normalization, data analysis, MIQE compliant qPCR workflow control, digital PCR applications and assays, MIQE compliance using qPCR arrays and much more. iBook has been written by renown experts in the field: Michael Pfaffl, Afif Abdel Nour, Stephen A. Bustin, Jim Huggett, Mikael Kubista, Jan Hellemans, Jo Vandesompele, Tania Nolan, Eva Schmidt etc. In collaboration with companies: Roche Diagnostics, Agilent Technologies, Gilson Inc., BioSistemika LLC, Bio-Rad, TATAA, Fluidigm etc.
We believe MIQE guidelines should be free for everyone, so the iBook is free. It is a ‘live’ and interactive iBook with 13 chapters, put together by 50 collaborators. The book contains maps, pictures and quizzes. It contains references that will direct you to PubMed and other relevant websites. We want to keep it easy to learn, fun to use and easy to share and quote. The book will be regularly upgraded with new relevant information.
Dr Afif Abdel Nour, Field Application Scientist, Bio-Rad Laboratories
The MIQE guidelines’ popularity is growing within the global research community. Software applications (for smartphones, tablets and PCs) have been downloaded thousands of times. More information on the MIQE software applications is available here: Abdel Nour et al. 2013
For more relevant information please visit MIQE Gene Quantification Info Website.
Michael W. Pfaffl (TUM Munich), Klemen Zupancic (BioSistemika), Jim Huggett (LGC Group), Stephen A. Bustin (Anglia Ruskin University) at the excellent qPCR & NGS Event, Freising-Weihenstephan, Germany 2015.
By Tea Pavlek, BioSistemika LLC