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dc.contributor.author
Dionisi, Hebe Monica  
dc.contributor.author
Harms, Gerda  
dc.contributor.author
Layton, Alice C.  
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Gregory, Igrid R.  
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Parker, Jack J.  
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Hawkins, Shawn A.  
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Robinson, Kevin G.  
dc.contributor.author
Sayler, Gary S.  
dc.date.available
2020-05-06T14:44:42Z  
dc.date.issued
2003-11  
dc.identifier.citation
Dionisi, Hebe Monica; Harms, Gerda; Layton, Alice C.; Gregory, Igrid R.; Parker, Jack J.; et al.; Power analysis for real-time PCR quantification of genes in activated sludge and analysis of the variability introduced by DNA extraction; American Society for Microbiology; Applied And Environmental Microbiology; 69; 11; 11-2003; 6597-6604  
dc.identifier.issn
0099-2240  
dc.identifier.uri
http://hdl.handle.net/11336/104320  
dc.description.abstract
The aims of this study were to determine the power of discrimination of the real-time PCR assay for monitoring fluctuations in microbial populations within activated sludge and to identify sample processing points where methodological changes are needed to minimize the variability in target quantification.  DNA was extracted using a commercially available kit from  mixed liquor samples taken from the aeration tank of four bench-scale activated-sludge reactors operating at 2-, 5-, 10- and 20-day solids retention times, with mixed-liquor volatile suspended solid (MLVSS) values ranging from 260 to 2,610 mg/L. Real-time PCR assays for bacterial and Nitrospira 16S rRNA genes were chosen because they represent, respectively, a highly abundant and a lower abundant bacterial target subject to clustering within the activated sludge matrix. The mean coefficient of variation in DNA yields (measured as µg DNA per mg MLVSS) in triplicate extractions of twelve different samples was 12.2%.  Based on power analyses, the variability associated with DNA extraction had a small impact on the overall variability of the real-time PCR assay.   Instead, a larger variability was associated with the PCR assay.   The less abundant target (Nitrospira 16S rRNA gene) had more variability than the highly abundant target (bacterial 16S rRNA gene) and samples from the lower biomass reactors had more variability than samples from the higher biomass reactors.   Power analysis of real-time PCR assays indicated that 3 to 5 samples were necessary to detect a 2-fold increase in bacterial 16S rRNA genes, whereas 3 to 5 samples were required to detect a 5-fold increase in Nitrospira 16S rRNA genes.  
dc.format
application/pdf  
dc.language.iso
eng  
dc.publisher
American Society for Microbiology  
dc.rights
info:eu-repo/semantics/openAccess  
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/  
dc.subject
BACTERIAL 16S rRNA GENE  
dc.subject
MIXED-LIQUOR  
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NITROSPIRA  
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POWER ANALYSIS  
dc.subject.classification
Biología Celular, Microbiología  
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Ciencias Biológicas  
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CIENCIAS NATURALES Y EXACTAS  
dc.title
Power analysis for real-time PCR quantification of genes in activated sludge and analysis of the variability introduced by DNA extraction  
dc.type
info:eu-repo/semantics/article  
dc.type
info:ar-repo/semantics/artículo  
dc.type
info:eu-repo/semantics/publishedVersion  
dc.date.updated
2020-04-06T16:10:17Z  
dc.identifier.eissn
1098-5336  
dc.journal.volume
69  
dc.journal.number
11  
dc.journal.pagination
6597-6604  
dc.journal.pais
Estados Unidos  
dc.conicet.avisoEditorial
Starting in 2016, Open access articles appearing in ASM journals are published under a Creative Commons Attribution 4.0 International license. The author(s) retains copyright under this license. Others may copy, distribute, adapt, reorganize, and build upon the published work, as long as credit to the author and original article is given, and the new work, which includes the previously published content, is licensed under identical terms. (Note that open access articles published prior to 2016 are covered under a variety of licenses and so we encourage users to review the license information carefully in order to determine how the content may be reused).  
dc.description.fil
Fil: Dionisi, Hebe Monica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico; Argentina  
dc.description.fil
Fil: Harms, Gerda. University of Tennessee; Estados Unidos  
dc.description.fil
Fil: Layton, Alice C.. University of Tennessee; Estados Unidos  
dc.description.fil
Fil: Gregory, Igrid R.. University of Tennessee; Estados Unidos  
dc.description.fil
Fil: Parker, Jack J.. University of Tennessee; Estados Unidos  
dc.description.fil
Fil: Hawkins, Shawn A.. University of Tennessee; Estados Unidos  
dc.description.fil
Fil: Robinson, Kevin G.. University of Tennessee; Estados Unidos  
dc.description.fil
Fil: Sayler, Gary S.. University of Tennessee; Estados Unidos  
dc.journal.title
Applied And Environmental Microbiology  
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://aem.asm.org/content/69/11/6597  
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://aem.asm.org/content/aem/69/11/6597.full.pdf  
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1128/AEM.69.11.6597-6604.2003