Research Output
Quantification of Ammonia Oxidizing Bacterial Abundances in Environmental Samples by Quantitative-PCR
  Quantitative-PCR (qPCR) enables the quantification of specific DNA targets, such as functional or phylogenetic marker genes associated with environmental samples. During each qPCR cycle, the number of copies of a gene (or region) of interest in DNA samples is determined in real time using a fluorescence-based label and compared to a standard serial dilution. Here, we describe a qPCR method to quantify the ammonia oxidizing bacteria involved in the first step of nitrification, using the amoA gene as a proxy of their abundance. The preparation of the standards from environmental samples and qPCR is presented in detail for specifically quantifying microbial abundance in environmental samples such as soil.

  • Date:

    08 November 2020

  • Publication Status:

    Published

  • Publisher

    Springer

  • DOI:

    10.1007/978-1-0716-1040-4_12

  • Cross Ref:

    10.1007/978-1-0716-1040-4_12

  • Funders:

    Edinburgh Napier Funded

Citation

Blaud, A., Maïder, A., & Clark, I. M. (2021). Quantification of Ammonia Oxidizing Bacterial Abundances in Environmental Samples by Quantitative-PCR. In L. C. Carvalhais, & P. G. Dennis (Eds.), The Plant Microbiome: Methods and Protocols (135-146). New York: Springer. https://doi.org/10.1007/978-1-0716-1040-4_12

Authors

Keywords

qPCR, PCR, Standard curve, Melting curve, Gel extraction, SYBR Green, Ammonia-oxidizing bacteria, amoA

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