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T-RFLP Analysis

A Rapid Fingerprinting Method for Studying Diversity, Structure, and Dynamics of Microbial Communities

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Environmental Microbiology

Part of the book series: Methods in Biotechnology ((MIBT,volume 16))

Abstract

Terminal restriction fragment length polymorphism (T-RFLP) analysis is a method for rapid profiling of mixed populations of an homologous amplicon (i.e., diverse sequences of a single gene). It combines restriction fragment analysis of a PCR-amplified gene marker with automated sequencing gel technology. One primer used in PCR amplification of the marker gene is labeled at the 5′ terminus with a fluorescent dye, in order that the terminal restriction fragments (T-RFs) of the digested amplicon can be detected and quantified (1–3). Detailed evaluations of T-RFLP analysis have shown that, in most cases, both the sizes and relative signal intensities of the individual T-RFs in a sample are highly reproducible. Consequently, T-RFLP analysis is an excellent tool for rapidly comparing microbial communities (4–6).

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Authors and Affiliations

  1. Max-Planck-Institute for Terrestrial Microbiology, Marburg, Germany

    Werner Liesack & Peter F. Dunfield

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  1. Werner Liesack
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  2. Peter F. Dunfield
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Editors and Affiliations

  1. Planta Piloto de Procesos Industriales Microbiológicos (PROIMI)-CONICET, San Miguel de Tucumán, Argentina

    John F. T. Spencer  & Alicia L. Ragout de Spencer  & 

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Liesack, W., Dunfield, P.F. (2004). T-RFLP Analysis. In: Walker, J.M., Spencer, J.F.T., Ragout de Spencer, A.L. (eds) Environmental Microbiology. Methods in Biotechnology, vol 16. Humana Press. https://doi.org/10.1385/1-59259-765-3:023

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  • DOI: https://doi.org/10.1385/1-59259-765-3:023

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-116-5

  • Online ISBN: 978-1-59259-765-9

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