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

  • Töwe S
  • Kleineidam K
  • Schloter M
  • 92

    Readers

    Mendeley users who have this article in their library.
  • 28

    Citations

    Citations of this article.

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. © 2010 Elsevier B.V.

Author-supplied keywords

  • Amplification efficiency
  • Functional genes
  • LinRegPCR
  • Nitrogen cycle
  • Quantitative real-time PCR
  • Standard curve

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document

Authors

  • Stefanie Töwe

  • Kristina Kleineidam

  • Michael Schloter

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free