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Addressing fluorogenic real-time qpcr inhibition using the novel custom excel file system ‘FocusField2-6GallupqPCRSet-upTool-1’ to attain consistently high fidelity qPCR reactions

External protocol Created on 09 Apr 2014

Authors

Jack M. Gallup, Mark R. Ackermann

Summary

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.

Further details

The protocol was imported from Springer Protocols. To see the entire protocol, click on the source link, or download the pdf here.

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