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Molecular analysis of bacterial community DNA in sludge undergoing autothermal thermophilic aerobic digestion (ATAD): pitfalls and improved methodology to enhance diversity recovery

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dc.contributor.author Pembroke, Tony J.
dc.contributor.author Piterina, Anna Valentinova
dc.contributor.author Bartlett, John
dc.date.accessioned 2010-07-14T16:07:34Z
dc.date.available 2010-07-14T16:07:34Z
dc.date.issued 2010
dc.identifier.citation Piterina, Anna V.; Bartlett, John; Pembroke, J. Tony. 2010. "Molecular Analysis of Bacterial Community DNA in Sludge Undergoing Autothermal Thermophilic Aerobic Digestion (ATAD): Pitfalls and Improved Methodology to Enhance Diversity Recovery." Diversity 2(4), 505-526 en_US
dc.identifier.uri http://hdl.handle.net/10344/401
dc.description peer-reviewed en_US
dc.description.abstract Molecular analysis of the bacterial community structure associated with sludge processed by autothermal thermophilic aerobic digestion (ATAD), was performed using a number of extraction and amplification procedures which differed in yield, integrity, ability to amplify extracted templates and specificity in recovering species present. Interference to PCR and qPCR amplification was observed due to chelation, nuclease activity and the presence of thermolabile components derived from the ATAD sludge. Addition of selected adjuvant restored the ability to amplify community DNA, derived from the thermophilic sludge, via a number of primer sets of ecological importance and various DNA polymerases. Resolution of community profiles by molecular techniques was also influenced by the ATAD sludge extraction procedure as demonstrated by PCR-DGGE profiling and comparison of taxonomic affiliations of the most predominant members within 16S rRNA gene libraries constructed from ATAD DNA extracted by different methods. Several modifications have been shown to be necessary to optimize the molecular analysis of the ATAD thermal niche which may have general applicability to diversity recovery from similar environments. en_US
dc.language.iso eng en_US
dc.publisher MDPI en_US
dc.relation.ispartofseries Diversity;2/4/ p505-526
dc.subject ATAD en_US
dc.subject thermophilic sludge en_US
dc.subject DNA extraction en_US
dc.subject PCR optimisation en_US
dc.subject PCR inhibition en_US
dc.subject microbial community biodiversity en_US
dc.subject molecular analysis en_US
dc.subject clone library en_US
dc.title Molecular analysis of bacterial community DNA in sludge undergoing autothermal thermophilic aerobic digestion (ATAD): pitfalls and improved methodology to enhance diversity recovery en_US
dc.type Journal Article en_US
dc.type.supercollection all_ul_research en_US
dc.type.supercollection ul_published_reviewed en_US
dc.type.restriction none en
dc.contributor.sponsor HEA


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