Differences in amplification efficiency of standard curves in quantitative real-time PCR assays and consequences for gene quantification in environmental samples

Journal of Microbiological Methods
Stefanie TöweMichael Schloter

Abstract

High and comparable efficiency values are the key for reliable quantification of target genes from environmental samples using real-time PCR. Therefore it was the aim of this study to investigate if PCR amplification efficiencies of plasmid DNA used for the calculation of standard curves (i) remain constant along a logarithmic scale of dilutions and (ii) if these values are comparable to those of DNA extracted from environmental samples. It could be shown that comparable efficiency values within the standards cannot be achieved using log scale serial dilutions and a comparison of gene copy numbers from DNA extracted from environmental samples and standard DNA extracted from plasmids is only possible in a very small interval.

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Citations

Mar 28, 2012·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Ian M ClarkPenny R Hirsch
Apr 12, 2012·Applied and Environmental Microbiology·Robert BrankatschkHelmut Bürgmann
Aug 8, 2015·Journal of the Science of Food and Agriculture·Alessandro FlorioAnna Benedetti
Jan 26, 2017·Scientific Reports·Alexander NagyMartina Havlíčková
Jun 8, 2017·Environmental Monitoring and Assessment·Loredana CanforaRosa Francaviglia
Jan 21, 2015·Journal of Environmental Quality·Kristopher S GuentzelJessica L Kozarek
Apr 24, 2021·Clinical Chemistry·Jan M RuijterMaurice J B van den Hoff

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