By measuring the kinetics of the reaction in the early phases of the polymerase chain reaction (PCR), quantitative real time PCR (qRT-PCR) allows the rapid detection and quantification of nucleic acids. As part of BioMed Central’s ongoing efforts to promote data standards and encourage the normalization and optimization of widely used technologies – as exemplified by the BMC Research Notes series on microarrays – BMC Research Notes is launching a new topical series on quantitative real-time PCR normalization and optimization.
qRT-PCR is a commonly used technology with a wide range of applications such as viral quantitation, pathogen detection, quantification of gene expression and oligonucleotide array validation. But the validity and reproducibility of results of qRT-PCR studies depend on a number of factors including the appropriate selection and validation of reference genes, the sufficient reporting of experimental settings – details of which are listed in the MIQE guidelines (Minimum Information for Publication of Quantitative Real-Time PCR Experiments) and ultimately the use of validated statistical tools and methods of data analysis.
The series, edited by Dr Joshua Yuan Associate Professor in Bioinformatics at the Texas A&M University Department of Plant Pathology and Microbiology, brings together descriptions of reference genes [1, 2] and potential associated experimental artifacts as well as tools for the improvement of RT-PCR performance [3, 4] and data analysis.
In-house Editor – BMC Research Notes
MIQE has been a breath of fresh air for the qPCR world (note that only the term qPCR conforms with the guidelines to the exclusion of qRT-PCR). The MIQE guidelines have given credibility to a technique that suffered from lack of standardized practice. For a brief review of the guidelines in accordance with one of the authors of the Methods paper, check out A practical approach to MIQE for the bench scientist