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

Abstract

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.

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Correspondence to Jack M. Gallup.

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Open Access This article is published under license to BioMed Central Ltd. This is an Open Access article is distributed under the terms of the Creative Commons Attribution License ( https://creativecommons.org/licenses/by/2.0 ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Gallup, J.M., Ackermann, M.R. Addressing fluorogenic real-time qpcr inhibition using the novel custom excel file system ‘FocusField2-6GallupqPCRSet-upTool-001’ to attain consistently high fidelity qPCR reactions. Biol. Proced. Online 8, 87–153 (2006). https://doi.org/10.1251/bpo122

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  • DOI: https://doi.org/10.1251/bpo122

Indexing terms

  • Reverse Transcription
  • DNA, Complementary