Determining a minimum detection threshold in terminal restriction fragment length polymorphism analysis
dc.contributor.author | Courtney, Kevin C. | |
dc.contributor.author | Bainard, Luke D. | |
dc.contributor.author | Sikes, Benjamin A. | |
dc.contributor.author | Koch, Alexander M. | |
dc.contributor.author | Maherali, Hafiz | |
dc.contributor.author | Klironomos, John N. | |
dc.contributor.author | Hart, Miranda M. | |
dc.date.accessioned | 2015-11-05T21:30:11Z | |
dc.date.available | 2015-11-05T21:30:11Z | |
dc.date.issued | 2012-01 | |
dc.identifier.citation | KC Courtney†, LD Bainard, BA Sikes, AM Koch, MM Hart, H Maherali, JN Klironomos (2012) Determining a minimum detection threshold in terminal restriction fragment length polymorphism analysis. Journal of Microbiological Methods 88(1): 14-18. http://dx.doi.org/10.1016/j.mimet.2011.09.016 | en_US |
dc.identifier.uri | http://hdl.handle.net/1808/18838 | |
dc.description.abstract | Terminal restriction fragment length polymorphism (T-RFLP) analysis is a common technique used to characterize soil microbial diversity. The fidelity of this technique in accurately reporting diversity has not been thoroughly evaluated. Here we determine if rare fungal species can be reliably detected by T-RFLP analysis. Spores from three arbuscular mycorrhizal fungal species were each mixed at a range of concentrations (1%, 10%, 50%, and 100%) with Glomus irregulare to establish a minimum detection threshold. T-RFLP analysis was capable of detecting diagnostic peaks of rare taxa at concentrations as low as 1%. The relative proportion of the target taxa in the sample and DNA concentration influenced peak detection reliability. However, low concentrations produced small, inconsistent electropherogram peaks contributing to difficulty in differentiating true peaks from signal noise. The results of this experiment suggest T-RFLP is a reproducible and high fidelity procedure, which requires careful data interpretation in order to accurately characterize sample diversity. | en_US |
dc.publisher | Elsevier | en_US |
dc.rights | This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License 4.0 (CC BY-NC-ND 4.0), which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made. | |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/ | |
dc.subject | T-RFLP | en_US |
dc.subject | Detection threshold | en_US |
dc.subject | Arbuscular mycorrhizal | en_US |
dc.subject | Fungi | en_US |
dc.subject | Spores | en_US |
dc.title | Determining a minimum detection threshold in terminal restriction fragment length polymorphism analysis | en_US |
dc.type | Article | |
kusw.kuauthor | Sikes, Benjamin A. | |
kusw.kudepartment | Ecology & Evolutionary Biology | en_US |
dc.identifier.doi | 10.1016/j.mimet.2011.09.016 | |
kusw.oaversion | Scholarly/refereed, publisher version | |
kusw.oapolicy | This item meets KU Open Access policy criteria. | |
dc.rights.accessrights | openAccess |
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Except where otherwise noted, this item's license is described as: This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License 4.0 (CC BY-NC-ND 4.0), which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.