Abstract
Quantitative PCR is the “gold standard” technology to quantify nucleic acids and, since the first report describing real-time PCR detection in 1993, its use has been grown exponentially. More recent technological advancements have extended the field of applications ranging from high-resolution melting detection to digital PCR. Nowadays, it is a very accessible technique, but some pitfalls should be overcome in order to achieve robust and reliable analysis.
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References
Higuchi R, Fockler C, Dollinger G et al (1993) Kinetic PCR analysis: real-time monitoring of DNA amplification reactions. Biotechnology (NY) 11:1026–1030
Mullis KF, Faloona S, Scharf R et al (1986) Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction. Cold Spring Harbor Symp Quant Biol 51:263–273
Bustin SA, Nolan T (2004) Pitfalls of quantitative real-time reversetranscription polymerase chain reaction. Biomol Tech 15:155–166
Ishiguro T, Saitoh J, Yawata H et al (1995) Homogeneous quantitative assay of hepatitis C virus RNA by polymerase chain reaction in the presence of a fluorescent intercalater. Anal Biochem 229:207–213
Bonnet G, Tyagi S, Libchaber A et al (1999) Thermodynamic basis of the enhanced specificity of structured DNA probes. PNAS USA 96: 6171–6176
Bustin SA, Benes V, Garson JA et al (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55:611–622
Raymaekers M, Smets R, Maes B et al (2009) Checklist for optimization and validation of real-time PCR assays. J Clin Lab Anal 23: 145–151
Taylor S, Wakem M, Dijkman G et al (2010) A practical approach to RT-qPCR-Publishing data that conform to the MIQE guidelines. Methods 50:S1–S5
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Raso, A., Biassoni, R. (2014). Twenty Years of qPCR: A Mature Technology?. In: Biassoni, R., Raso, A. (eds) Quantitative Real-Time PCR. Methods in Molecular Biology, vol 1160. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0733-5_1
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DOI: https://doi.org/10.1007/978-1-4939-0733-5_1
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0732-8
Online ISBN: 978-1-4939-0733-5
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