In the last Newsletter we introduced the MIQE guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments, a set of standards originally published in 2009, then revised slightly in 2011.
The following Newsletter explains how thermo fisher’s large and diverse collection of TaqMan® assays and consumables comply with these guidelines.
ThermoFisher’s qPCR assays are designed using an advanced bioinformatics pipeline that ensures maximum amplification efficiency, standardized running conditions requiring no optimization, and 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. ThermoFisher’s performs regular remapping based on revisions to the NCBI databases, helping ensure that assays and their associated information remain current. When you search for an assay on the website, you 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. You can see the context sequence, the probe location, and the exon-exon boundary that the assay crosses. You can also consult a map showing how all assays for a given gene line up with all know transcripts. ThermoFisher provides all information necessary for choosing the right assay to meet experimental needs, and for meeting the requirements set out by MIQE. TaqMan Assays have been offered for many years, with over 5000 publications citing their use.
As mentioned in the MIQE guidelines:” because the consumables used affect thermal cycling, it is necessary to identify the use of single tubes, strips, or plates, and their manufacturers. The degree of transparency of the plasticware used, is also important, because different plastics exhibit substantial differences in fluorescence reflection and sensitivity. When plates are used, the method of sealing can affect the evaporation of samples at the plate”. Applied Biosystems MicroAmp plastics consumables offer excellent PCR or qPCR performance in formats developed to meet your experimental needs.