A Practical Approach to Assay Design for qPCR – Overcoming Difficult Assays, Designs and Optimizations while Conforming to the MIQE Guidelines

Francisco Bizouarn Bio-Rad Laboratories, United States of America

Abstract
Quantitative PCR assays in the past have often been run without serious specificity and quality considerations. Target sequence information was inserted into “black-box” software packages and assays run under general conditions. With the arrival of MIQE guidelines, a greater emphasis is placed on the use of robust and validated assays. Generating robust assays is generally a simple task, but occasionally obstacles can be encountered. Ease in the optimization of an assay is often directly linked to the original design. In this talk, we will present critical assay design parameters and tips to overcome potential problems, as well as quick optimization and validation strategies that help fulfil MIQE guidelines.