Real-time PCR probe optimization using design of experiments approach

16.12.2015

Wadle S, Lehnert M, Rubenwolf S, Zengerle R, von Stetten F

Biomolecular Detection and Quantification 7 (2016) 1-8

NBiomolecular Detection and Quantification            online article

Primer and probe sequence designs are among the most critical input factors in real-time polymerase chain reaction (PCR) assay optimization. In this study, we present the use of statistical design of experiments (DOE) approach as a general guideline for probe optimization and more specifically focus on design optimization of label-free hydrolysis probes that are designated as mediator probes (MPs), which are used in reverse transcription MP PCR (RT-MP PCR). The DOE approach produces excellent results and may also reduce the number of experiments, thus reducing costs and experimental times.