- Please read the THE MIQE GUIDELINES: Bustin, S.A., Benes, V., Garson, J.A., Hellemans, J., Huggett,
J., Kubista, M., Mueller, R., Nolan, T., Pfaffl, M.W., Shipley, G.L., Vandesompele, J., and Wittwer, C.T. (2009). The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments. Clin. Chem. 55, 611-622. Download.pdf
- Invited book chapter by J.M. Gallup for Horizon Scientific Press, “Difficult Templates and Inhibitors of PCR” in the book: “PCR Troubleshooting and Optimization: The Essential Guide” January 2011. Click here
- Gallup, J.M., Kawashima, K., Lucero, G., and Ackermann, M.R. (2005). New quick method for isolating RNA from laser captured cells stained by immunofluorescent immunohistochemistry; RNA suitable for direct use in fluorogenic TaqMan one-step real-time RT-PCR. Biol. Proced. Online. 7, 70-92. Download.pdf
- Gallup, J.M., and Ackermann M.R. (2006). 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, 87-155. Download.pdf
- Gallup, J.M., and Ackermann, M.R. (2008). The ‘PREXCEL-Q Method’ for qPCR. Int. J. Biomed. Sci. 4,
273-293. Download.pdf
- PubMed PDF of The ‘PREXCEL-Q Method’ for qPCR. Download.pdf
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Sponseller, B.A., de Macedo, M.M., Clark, S.K., Gallup, J.M., and Jones, D.E. (2009). Activation of
peripheral blood monocytes results in more robust production of IL-10 in neonatal foals compared
to adult horses. Vet. Immunol. Immunopathol. 127, 167-173. Download.pdf
- Sow, F.B., Gallup, J.M., Meyerholz, D.K., and Ackermann, M.R. (2009). Gene profiling studies in the neonatal ovine lung show enhancing effects of VEGF on the immune response. Dev. Comp. Immunol.
33, 761-771. Download.pdf
- Sow, F.B., Gallup, J.M., Sacco, R.E., and Ackermann, M.R. (2009). Laser Capture Microdissection Revisited as a Tool for Transcriptomic Analysis: Application of an Excel-Based qPCR Preparation
Software (PREXCEL-Q). Int. J. Biomed. Sci. 5, 105-124. Download.pdf
- Gallup, J.M., Sow, F.B., Van Geelen, A., and Ackermann, M.R. (2009). SPUD qPCR Assay Confirms PREXCEL-Q Software’s Ability to Avoid qPCR Inhibition. Curr. Issues Mol. Biol. 12, 129-134. Download.pdf
- Bustin, S.A. (2008). Real-time polymerase chain reaction – towards a more reliable, accurate and relevant assay. Eur. Pharm. Rev. 6, 19-27. [see p. 24, column 2, paragraph 3] Download.pdf
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Bustin, S.A. (2008). Real-time quantitative PCR – opportunities and pitfalls. Eur. Pharm. Rev. 4, 18-23.
Download.pdf -
Murphy, J., and Bustin, S.A. (2009). Reliability of real-time reverse transcription PCR in clinical diagnostics:
gold standard or substandard? Expert Rev. Mol. Diagn. 9(2), 187-197. Download.pdf -
Practical implementation of minimum standard (MIQE) guidelines for fluorescence-based real-time quantitative PCR experiments Download.pdf
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Stephen A Bustin, Vladimir Benes, Jeremy Garson, Jan Hellemans, Jim Huggett, Mikael Kubista, Reinhold Mueller, Tania Nolan, Michael W Pfaffl, Gregory Shipley, Jack M Gallup, Gert Van Peer, Mario Van Poucke, Nadine Van Roy, Sarah Vergult, Joris Wauman, Marina Tshuikina-Wiklander, Erik Willems, Sara Zaccara, Fjoralba Zeka, Jo Vandesompele, et al. The need for transparency and good practices in the qPCR literature. Nature Methods (Impact Factor: 23.57). 11/2013; 10(11):1063-1067. DOI: 10.1038/nmeth.2697. Download.pdf
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Other related links: Quanta BioSciences
- Join in the qPCR fun at BioTechniques Molecular Biology Techniques Forums
- In support of PREXCEL-Q
- Dr. Stephen A. Bustin's site, and the related RDML Consortium site
- Mikael Kubista of the TATAA Biocenter speaks on important aspects of good qPCR
- Gene Quantification Download site by Michael W. Pfaffl> Home site
- BIO-RAD ppt on MIQE workflow ideas
- Link to MO BIO Laboratories, Inc. (for RNA and DNA purification and inhibitor removal)
- Solaris "splice variant aware" primer-probe design (remember to address splice variants!)
- Some additional key suggestions for performing good qPCR
- On qPCR problems and inhibition
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For PREXCEL-Q (ISURF Software #03407) please contact Dr. Dario Valenzuela
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to request an End User License Agreement (EULA) from Iowa State University
Research Foundation (ISURF) at: dariov@iastate.edu
ISU Office of Biotechnology announcement
Click here to visit the DNA Facility of the Iowa State University Office of Biotechnology
qPCR service at the DNA Facility of the Iowa State University Office of Biotechnology