Scott D. Rose
Integrated DNA Technologies, United States of America
Well-designed qPCR assays require the careful consideration of primer placement, specificity, avoidance of single nucleotide polymorphisms, oligonucleotide interactions and accurate melting temperature calculations. Using pre-designed assays shifts the burden of assay development from the researcher to the manufacturer. Unfortunately, some of the pre-designed assay sets commercially available today were built using outdated databases and primer design was done with older, less accurate Tm algorithms. Even worse, some suppliers do not provide sequence information to the end user that would enable the researcher to evaluate the reagents in the context of their specific requirements or for use in publication. IDT now offers fully guaranteed PrimeTime® Pre-designed qPCR assays (for all genes in the human, mouse, and rat genome) that avoid cross reactivity within that genome, known single nucleotide polymorphisms, primer interactions, and gives full disclosure of all sequence information as recommended by MIQE guidelines. By including ZEN double-quenched probes, these assays offer sensitivity to 10 copies or lower with the added security of knowing precise probe location with full sequence disclosure.
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