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dc.contributor.author
Dionisi, Hebe Monica
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dc.contributor.author
Harms, Gerda
dc.contributor.author
Layton, Alice C.
dc.contributor.author
Gregory, Igrid R.
dc.contributor.author
Parker, Jack J.
dc.contributor.author
Hawkins, Shawn A.
dc.contributor.author
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
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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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dc.subject.classification
Ciencias Biológicas
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dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
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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
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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
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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
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