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Biological Procedures Online

Open Access

Addressing fluorogenic real-time qpcr inhibition using the novel custom excel file system ‘FocusField2-6GallupqPCRSet-upTool-001’ to attain consistently high fidelity qPCR reactions

Biological Procedures Online8:81087

Received: 2 May 2006

Accepted: 10 July 2006


The purpose of this manuscript is to discuss fluorogenic real-time quantitative polymerase chain reaction (qPCR) inhibition and to introduce/define a novel Microsoft Excel-based file system which provides a way to detect and avoid inhibition, and enables investigators to consistently design dynamically-sound, truly LOG-linear qPCR reactions very quickly. The qPCR problems this invention solves are universal to all qPCR reactions, and it performs all necessary qPCR set-up calculations in about 52 seconds (using a pentium 4 processor) for up to seven qPCR targets and seventytwo samples at a time - calculations that commonly take capable investigators days to finish. We have named this custom Excel-based file system “FocusField2-6GallupqPCRSet-upTool-001” (FF2-6-001 qPCR set-up tool), and are in the process of transforming it into professional qPCR set-up software to be made available in 2007. The current prototype is already fully functional.

Indexing terms

Reverse TranscriptionDNA, Complementary