Following the MIQE Guidelines for RT qPCR Experiments

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Researchers -- and journals -- have been slow to adopt the MIQE guidelines that were established in 2009 to bolster the reliability of real-time PCR (qPCR) and reverse transcription qPCR (RT-qPCR) data. In response, Sean Taylor and Eli Mrkusich wrote a brief and practical guide that concisely summarizes the key steps required to produce high quality, reproducible data for labs conducting RT-qPCR experiments. This video highlights the key steps required to produce reliable and reproducible qPCR data as described in an article, published in the Journal of Molecular Microbiology and Biotechnology in November, 2013, by Bio-Rad scientists Sean Taylor and Eli Mrkusic (see http://www.bio-rad.com/en-us/life-sci....

Bio-Rad's PrimePCR™ PCR Primers
Bio-Rad collaborated with Biogazelle, leaders in real-time PCR research, to design and experimentally validate PCR primers for gene expression assays across the human and mouse transcriptomes. All PCR primers were designed to meet stringent performance standards following the MIQE guidelines (Minimum information for publication of quantitative real-time PCR experiments; Bustin et al. 2009).

These DNA primer pairs were designed by prioritizing the gene regions most commonly found in transcript variants. Strict design criteria were used to ensure optimal real-time PCR results for each target:
• Target regions without SNPs
• PCR primer pairs annealing across intron/exon junctions when possible
• No secondary structure in primer annealing sites
• Maximum number of transcript isoforms detected
• PCR primers compatible with standard assay conditions

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