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Open Access Green as soon as Postprint is submitted to ZB.
Differences in amplification efficiency of standard curves in quantitative real-time PCR assays and consequences for gene quantification in environmental samples.
J. Microbiol. Methods 82, 338-341 (2010)
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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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Quantitative real-time PCR; Amplification efficiency; Standard curve; LinRegPCR; Nitrogen cycle; Functional genes
ISSN (print) / ISBN
0167-7012
e-ISSN
1872-8359
Quellenangaben
Volume: 82,
Issue: 3,
Pages: 338-341
Publisher
Elsevier
Publishing Place
Amsterdam
Reviewing status
Peer reviewed
Institute(s)
Institute of Soil Ecology (IBOE)