期刊
JOURNAL OF MICROBIOLOGICAL METHODS
卷 82, 期 3, 页码 338-341出版社
ELSEVIER
DOI: 10.1016/j.mimet.2010.07.005
关键词
Quantitative real-time PCR; Amplification efficiency; Standard curve; LinRegPCR; Nitrogen cycle; Functional genes
资金
- Deutsche Forschungsgemeinschaft (DFG, Bonn) [SFB/TRR38]
- Brandenburg Ministry of Science, Research and Culture (MWFK, Potsdam)
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. (C) 2010 Elsevier B.V. All rights reserved.
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