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Article: Development of Quantitative Real-time PCR Assays for Different Clades of “Candidatus Accumulibacter”

TitleDevelopment of Quantitative Real-time PCR Assays for Different Clades of “Candidatus Accumulibacter”
Authors
Issue Date2016
PublisherNature Research: Fully open access journals. The Journal's web site is located at http://www.nature.com/srep/index.html
Citation
Scientific Reports, 2016, v. 6 n. 1, p. article no. 23993 How to Cite?
AbstractWe designed novel quantitative real-time polymerase chain reaction (qPCR) primers for the polyphosphate kinase 1 (ppk1) gene, targeting eight individual “Candidatus Accumulibacter” (referred to as Accumulibacter) clades. An evaluation of primer sets was conducted regarding the coverage, specificity, and PCR efficiency. (i) All primer sets were designed to cover all available sequences of the target clade. (ii) The phylogenetic analysis of the sequences retrieved from the qPCR products by each primer set demonstrated a high level of specificity. (iii) All calibration curves presented high PCR efficiencies in the range of 85–112% (R2 = 0.962–0.998). In addition, the possible interference of non-target amplicons was individually examined using the qPCR assay for 13 Accumulibacter clades, which were either undetected or showed negligible detection. With the primers designed by other research groups, a highly selective and sensitive qPCR-based method was developed to quantify all Accumulibacter clades, with the exception of Clade IE, in one assay, which enables more comprehensive insights into the community dynamics. The applicability to environmental samples was demonstrated by profiling the Accumulibacter clades in activated sludge samples of nine full-scale wastewater treatment plants.
Persistent Identifierhttp://hdl.handle.net/10722/293315
ISSN
2023 Impact Factor: 3.8
2023 SCImago Journal Rankings: 0.900
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorZhang, AN-
dc.contributor.authorMao, Y-
dc.contributor.authorZhang, T-
dc.date.accessioned2020-11-23T08:14:59Z-
dc.date.available2020-11-23T08:14:59Z-
dc.date.issued2016-
dc.identifier.citationScientific Reports, 2016, v. 6 n. 1, p. article no. 23993-
dc.identifier.issn2045-2322-
dc.identifier.urihttp://hdl.handle.net/10722/293315-
dc.description.abstractWe designed novel quantitative real-time polymerase chain reaction (qPCR) primers for the polyphosphate kinase 1 (ppk1) gene, targeting eight individual “Candidatus Accumulibacter” (referred to as Accumulibacter) clades. An evaluation of primer sets was conducted regarding the coverage, specificity, and PCR efficiency. (i) All primer sets were designed to cover all available sequences of the target clade. (ii) The phylogenetic analysis of the sequences retrieved from the qPCR products by each primer set demonstrated a high level of specificity. (iii) All calibration curves presented high PCR efficiencies in the range of 85–112% (R2 = 0.962–0.998). In addition, the possible interference of non-target amplicons was individually examined using the qPCR assay for 13 Accumulibacter clades, which were either undetected or showed negligible detection. With the primers designed by other research groups, a highly selective and sensitive qPCR-based method was developed to quantify all Accumulibacter clades, with the exception of Clade IE, in one assay, which enables more comprehensive insights into the community dynamics. The applicability to environmental samples was demonstrated by profiling the Accumulibacter clades in activated sludge samples of nine full-scale wastewater treatment plants.-
dc.languageeng-
dc.publisherNature Research: Fully open access journals. The Journal's web site is located at http://www.nature.com/srep/index.html-
dc.relation.ispartofScientific Reports-
dc.rightsScientific Reports. Copyright © Nature Research: Fully open access journals.-
dc.rightsThis work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.-
dc.titleDevelopment of Quantitative Real-time PCR Assays for Different Clades of “Candidatus Accumulibacter”-
dc.typeArticle-
dc.identifier.emailZhang, T: zhangt@hkucc.hku.hk-
dc.identifier.authorityZhang, T=rp00211-
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.1038/srep23993-
dc.identifier.scopuseid_2-s2.0-84966349216-
dc.identifier.hkuros319459-
dc.identifier.volume6-
dc.identifier.issue1-
dc.identifier.spagearticle no. 23993-
dc.identifier.epagearticle no. 23993-
dc.identifier.isiWOS:000375395500001-
dc.publisher.placeUnited Kingdom-
dc.identifier.issnl2045-2322-

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