Mittwoch, 12. August 2020

Relative expression software tool (REST©) for group-wise comparison and statistical analysis of relative expression results in real-time PCR

Relative Expression Software Tool (REST©) for group wise comparison
and statistical analysis of relative expression results in real-time PCR
Michael W. Pfaffl, Graham W. Horgan & Leo Dempfle (2002) 
Nucleic Acids Research 2002 30(9): E36

Real-time reverse transcription followed by polymerase chain reaction (RT–PCR) is the most suitable method for the detection and quantification of mRNA. It offers high sensitivity, good reproducibility and a wide quantification range. Today, relative expression is increasingly used, where the expression of a target gene is standardised by a non-regulated reference gene. Several mathematical algorithms have been developed to compute an expression ratio, based on real-time PCR efficiency and the crossing point deviation of an unknown sample versus a control. But all published equations and available models for the calculation of relative expression ratio allow only for the determination of a single transcription difference between one control and one sample. Therefore a new software tool was established, named REST© (relative expression software tool), which compares two groups, with up to 16 data points in a sample and 16 in a control group, for reference and up to four target genes. The mathematical model used is based on the PCR efficiencies and the mean crossing point deviation between the sample and control group. Subsequently, the expression ratio results of the four investigated transcripts are tested for significance by a randomisation test. Herein, development and application of REST© is explained and the usefulness of relative expression in real-time PCR using REST© is discussed. The latest software version of REST© and examples for the correct use can be downloaded at

Montag, 10. August 2020

Virtual Symposium -- ‪Diagnosing SARS-CoV-2 with qPCR‬

‪Diagnosing SARS-CoV-2 with qPCR‬

This virtual symposium, brought to you by The Scientist, will explore the limitations and challenges of current SARS-CoV-2 qPCR-based testing approaches, their root causes, and what can be learned for the future.
Tuesday, September 8, 2020
11:00 AM - 2:00 PM, Eastern Standard Time

Stephen A. Bustin, PhD
Professor of Molecular Medicine
Faculty of Health, Education, Medicine and Social Care
Anglia Ruskin University

Jim F. Huggett, PhD
Senior Lecturer, Analytical Microbiology
School of Biosciences and Medicine
University of Surrey

Mikael Kubista, PhD
Head, Institute of Biotechnology
Czech Academy of Sciences
Founder and Chairman of the Board
TATAA Biocenter

Jo Vandesompele, PhD
Department of Biomolecular Medicine
Faculty of Medicine and Health Sciences
Ghent University

Michael W. Pfaffl, PhD
Professor of Molecular Physiology
Animal Physiology & Immunology
School of Life Sciences
Technical University of Munich


Freitag, 31. Juli 2020

MicroGenomics 2021 -- September 29th till October 1st in Florence

Microgenomics 2021 will be hosting a workshop on September 29th, 2021 at the University of Florence, Italy. We look forward to seeing you there!
And REMEMBER, The symposium is postponed to September 30-October 1, 2021!

Dienstag, 14. Juli 2020

Our MIQE Guidelines show more than 10,000 citations

Congratulations to all authors!
Our MIQE Guidelines show more than 10,000 citations on Google Scholar
The MIQE Guidelines -- Minimum Information for Publication of Quantitative Real-Time PCR Experiments
#MIQE #qPCR #realtimePCR