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have been demonstrated in less than 10 minutes. Rapid cycling provides several benefits, including, reduced time to result, increased system throughput and improved reaction specificity. In practice however, engineering trade-offs between ease of use, temperature uniformity, and speed, mean that
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The noise in fluorescence measurements affects the precision of qPCR. It is typically a function of excitation source intensity variation, detector noise and mechanical noise. Multi factorial analysis has suggested that the contribution of mechanical noise is the most important factor, and that
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By using an instrument with sufficient optical channels and extensive assay optimisation, up to 7 separate targets can be simultaneously quantified in a single PCR reaction. However, even with extensive optimisation, the effective dynamic range of such multiplex assays is often reduced due to
62:
Thermal non-uniformity during temperature cycling contributes to variability in PCR and, unfortunately, some thermocyclers do not meet the specifications claimed by manufacturers. Increasing the speed of thermal cycling generally reduces thermal uniformity, and can reduce the
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The first quantitative PCR machine was described in 1993, and two commercial models became available in 1996. By 2009, eighteen different models were offered by seven different manufacturers. Prices range from about 4,300 USD to 150,000 USD
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The range of different fluorescent labels that can be monitored, the precision with which they can be measured, and the ability to discriminate signals from different labels, are relevant performance characteristics.
626:
Köppel, R.; Zimmerli, F.; Breitenmoser, A. (2009), "Heptaplex real-time PCR for the identification and quantification of DNA from beef, pork, chicken, turkey, horse meat, sheep (mutton) and goat",
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Schoder, D.; Schmalwieser, A.; Schauberger, G.; Hoorfar, J.; Kuhn, M.; Wagner, M. (2005), "Novel approach for assessing performance of PCR cyclers used for diagnostic testing",
74:
The temperature uniformity also has a direct effect on the ability to discriminate different PCR products by performing melting point analysis. In addition to uniformity, the
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Bahrdt, C.; Krech, A.; Wurz, A.; Wulff, D. (2010). "Validation of a newly developed hexaplex real-time PCR assay for screening for presence of GMOs in food, feed and seed".
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Hilscher, C.; Vahrson, W.; Dittmer, D.P. (2005), "Faster quantitative real-time PCR protocols may lose sensitivity and show increased variability",
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Therefore, the number of optical channels and the level of noise in fluorescence measurements are also important performance characteristics of
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Schoder, D.; Schmalwieser, A.; Schauberger, G.; Kuhn, M.; Hoorfar, J.; Wagner, M. (2003), "Physical characteristics of six new thermocyclers",
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Therefore, speed, precision and uniformity of thermal control are important performance characteristics of quantitative PCR instruments.
259:"Thermal analysis of the vortex tube based thermocycler for fast DNA amplification: Experimental and two-dimensional numerical results"
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Kim, Y.H.; Yang, I.; Bae, Y.S.; Park, S.R. (2008), "Performance evaluation of thermal cyclers for PCR in a rapid cycling condition",
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Higuchi, R.; Dollinger, G.; Walsh, P.S.; Griffith, R. (1992), "Simultaneous amplification and detection of specific DNA sequences",
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396:"Real-time PCR Machine System Modeling and a Systematic Approach for the Robust Design of a Real-time PCR-on-a-Chip System"
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Ma, H.; Shieh, K.; Chen, G.; Chen, X.; Chuang, M. (2006), "Application of Real-time
Polymerase Chain Reaction (RT-PCR)",
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Principal performance dimensions of quantitative PCR instruments are thermal control, fluorimetry and sample throughput.
587:"Amplicon melting analysis with labeled primers: a closed-tube method for differentiating homozygotes and heterozygotes"
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with which instruments are able to control temperature is a factor which affects their performance when performing
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Quantitative PCR instruments monitor the progress of PCR, and the nature of amplified products, by measuring
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systems with no moving parts in their optical paths are likely to provide improved quantitative precision.
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Wittwer, C.T.; Garling, D.J. (1991), "Rapid cycle DNA amplification: time and temperature optimization",
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Logan, J.; Edwards, K. (January 2009). "Chapter 2 An
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