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Do TaqMan® Assays Comply with MIQE Guidelines? -- Ask TaqMan® Ep. 19 by Life Technologies

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Published on Feb 25, 2013

Submit your real-time PCR questions at http://www.lifetechnologies.com/askta...
After introducing MIQE guidelines, a set of standards for producing and publishing qPCR data, Sr. Field Applications Specialist Doug Rains explains how Life Technologies' large and diverse collection of TaqMan® assays comply with these guidelines. Learn about Life Technologies' TaqMan assays guarantee program and the rigorous bioinformatics process through which each TaqMan® assays is designed.
Welcome to Ask TaqMan, where we answer your questions about real-time PCR. A concern among some researchers in the qPCR community is ensuring that experimental designs and data reporting practices meet the standards set out by MIQE Guidelines. Specifically, users often wonder whether or not Life Technologies' pre-developed TaqMan Assays comply with MIQE's set of recommendations. I'm here today to tell you that they do, and to explain how Life Technologies qPCR assays can help you to achieve the highest level of confidence in your data.
MIQE, which stands for Minimum Information for Publication of Quantitative Real-Time PCR Experiments, is a set of guidelines originally published in 2009, then revised slightly in 2011. The authors behind MIQE propose guidelines for all aspects of qPCR experimental design and analysis, as well as standards for sharing this information with colleagues.
So how do Life Tech's pre-developed qPCR Assays help labs fulfill MIQE recommendations with respect to design and information sharing? I'm glad you ask.
As explained both in the Life Technologies TaqMan Assays Guarantee Program white paper, all of Life Tech's million-plus qPCR assays are designed using an advanced bioinformatics pipeline that ensures maximum amplification efficiency, standardized running conditions requiring no optimization, and -- perhaps most importantly -- target specificity. During the design process, high-quality RefSeq context sequences are subjected to rigorous bioinformatic scrutiny using long-established design algorithms. This process produces gene-centric assays that won't amplify pseudogenes and homologues, while simultaneously avoiding known polymorphic sites and low-complexity sequences. What's more, Life technologies performs regular remapping based on revisions to the NCBI databases, helping ensure that assays and their associated information remain current.
When researchers search for an assay on our website, they can peruse a host of helpful information, including the NCBI gene ID, the gene symbol, a list of RefSeq IDs that are interrogated by that assay, and RNA accession numbers. And while we don't share the specific oligo sequences, we provide the context sequence, the probe location, and -- if appropriate -- the exon-exon boundary that the assay crosses. Finally, users can consult an a map showing how all assays for a given gene line up with all know transcripts.
In short, Life Technologies provides all information necessary for choosing just the right assay to meet experimental needs, and for meeting the requirements set out by MIQE. And best of all, TaqMan Assays have been offered for many years now, with well over 5000 publications citing their use.
If you'd like to learn more about searching for and selecting a TaqMan Assay that will suit your needs, please have a look at the Ask TaqMan video on this very topic.

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