Posted: November 07, 2013, by Sean Taylor
RT-qPCR (reverse transcription-quantitative polymerase chain reaction) is used routinely by a broad spectrum of researchers to assess the absolute or relative quantity of genomic DNA (gDNA) or complementary DNA (cDNA) in or between samples. Unfortunately, the many steps required for a successful experimental outcome have resulted in the adoption of a wide variation in methodologies, even within the same lab. This has led to publication of potentially misleading and even artifactual results that can slow the pace of research. The publication of the MIQE (Minimum Information for the publication of Quantitative real-time PCR Experiments) guidelines in 2009 and subsequent articles provided the scientific community with criteria to help achieve high-quality RT-qPCR results with valid conclusions. However, the majority of labs favor using their historical methods of experimentation over these highly recommended best practices. In an attempt to help remedy this ongoing challenge of adoption, a simplified roadmap based on the MIQE guidelines has been produced.