title: Influence of DNA isolation method on the investigation of archaeal diversity and abundance in biogas plants.
abstract: Various methods are available for DNA isolation from environmental samples. Because the chemical and biological composition of samples such as soil, sludge, or plant material is different, the effectiveness of DNA isolation can vary depending on the method applied and thus, have a substantial effect on the results of downstream analysis of the microbial community. Although the process of biogas formation is being intensely investigated, a systematic evaluation of kits for DNA isolation from material of biogas plants is still lacking. Since no DNA isolation kit specifically tailored for DNA isolation from sludge of biogas plants is available, this study compares five commercially available kits regarding their influence on downstream analyses such denaturing gradient gel electrophoresis (DGGE) and quantitative real-time PCR (qPCR). The results show that not all kits are equally suited for the DNA isolation from samples of different biogas plants, but highly reproducible DGGE fingerprints as well as qPCR results across the tested samples from biogas reactors using different substrate compositions could be produced using selected kits.
authors.affiliation: Sächsische Akademie der Wissenschaften zu Leipzig, Karl-Tauchnitz-Straße 1, 04107, Leipzig, Germany.
authors.forename: Juliane
authors.initials: J
authors.lastname: Theiss
authors.affiliation: Technische Universität Dresden, Professur für Mikrobielle Diversität, Zellescher Weg 20b, 01217, Dresden, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Rother
authors.affiliation: Sächsische Akademie der Wissenschaften zu Leipzig, Karl-Tauchnitz-Straße 1, 04107, Leipzig, Germany. kerstin.roeske@web.de.
authors.forename: Kerstin
authors.initials: K
authors.lastname: Röske
mesh_terms: D001105:Archaea / Q000235:genetics* / Q000378:metabolism; D044822:Biodiversity; D056804:Biofuels / Q000382:microbiology*; D019641:DNA, Archaeal / Q000235:genetics / Q000302:isolation & purification*; D058645:Denaturing Gradient Gel Electrophoresis / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012722:Sewage / Q000382:microbiology*; D012988:Soil Microbiology
chemical_list: D056804:Biofuels; D019641:DNA, Archaeal; D012336:RNA, Ribosomal, 16S; D012722:Sewage
keywords: Anaerobic digestion; DGGE; DNA isolation; Methanogenic Archaea; qPCR