caa a22 4500 605498423 CHVBK 20210128100546.0 cr unu---uuuuu 210128e20150801xx s 000 0 eng 10.1007/s00253-015-6721-z doi (NATIONALLICENCE)springer-10.1007/s00253-015-6721-z Design and application of a synthetic DNA standard for real-time PCR analysis of microbial communities in a biogas digester [Elektronische Daten] [T. May, M. Koch-Singenstreu, J. Ebling, R. Stantscheff, L. Müller, F. Jacobi, D. Polag, F. Keppler, H. König] A synthetic DNA fragment containing primer binding sites for the quantification of ten different microbial groups was constructed and evaluated as a reliable enumeration standard for quantitative real-time PCR (qPCR) analyses. This approach has been exemplary verified for the quantification of several methanogenic orders and families in a series of samples drawn from a mesophilic biogas plant. Furthermore, the total amounts of bacteria as well as the number of sulfate-reducing and propionic acid bacteria as potential methanogenic interaction partners were successfully determined. The obtained results indicated a highly dynamic microbial community structure which was distinctly affected by the organic loading rate, the substrate selection, and the amount of free volatile fatty acids in the fermenter. Methanosarcinales was the most predominant methanogenic order during the 3months of observation despite fluctuating process conditions. During all trials, the modified quantification standard indicated a maximum of reproducibility and efficiency, enabling this method to open up a wide range of novel application options. Springer-Verlag Berlin Heidelberg, 2015 qPCR nationallicence Microbial diversity nationallicence Methanogenic archaea nationallicence Anaerobic digester nationallicence Biogas plants nationallicence May T. Institute of Microbiology and Wine Research, Johannes Gutenberg University Mainz, Johann-Joachim Becherweg 15, 55128, Mainz, Germany aut Koch-Singenstreu M. Institute of Microbiology and Wine Research, Johannes Gutenberg University Mainz, Johann-Joachim Becherweg 15, 55128, Mainz, Germany aut Ebling J. Institute of Microbiology and Wine Research, Johannes Gutenberg University Mainz, Johann-Joachim Becherweg 15, 55128, Mainz, Germany aut Stantscheff R. Institute of Microbiology and Wine Research, Johannes Gutenberg University Mainz, Johann-Joachim Becherweg 15, 55128, Mainz, Germany aut Müller L. Deutsches Biomasseforschungszentrum gemeinnützige GmbH (DBFZ), Torgauer Straße 116, 04347, Leipzig, Germany aut Jacobi F. Deutsches Biomasseforschungszentrum gemeinnützige GmbH (DBFZ), Torgauer Straße 116, 04347, Leipzig, Germany aut Polag D. Institute of Earthsciences, Ruprecht-Karls University Heidelberg, Im Neuenheimer Feld 234-236, 69120, Heidelberg, Germany aut Keppler F. Institute of Earthsciences, Ruprecht-Karls University Heidelberg, Im Neuenheimer Feld 234-236, 69120, Heidelberg, Germany aut König H. Institute of Microbiology and Wine Research, Johannes Gutenberg University Mainz, Johann-Joachim Becherweg 15, 55128, Mainz, Germany aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/16(2015-08-01), 6855-6863 0175-7598 99:16<6855 2015 99 253 https://doi.org/10.1007/s00253-015-6721-z text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6721-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- May T. Institute of Microbiology and Wine Research, Johannes Gutenberg University Mainz, Johann-Joachim Becherweg 15, 55128, Mainz, Germany aut NATIONALLICENCE 700 1- Koch-Singenstreu M. Institute of Microbiology and Wine Research, Johannes Gutenberg University Mainz, Johann-Joachim Becherweg 15, 55128, Mainz, Germany aut NATIONALLICENCE 700 1- Ebling J. Institute of Microbiology and Wine Research, Johannes Gutenberg University Mainz, Johann-Joachim Becherweg 15, 55128, Mainz, Germany aut NATIONALLICENCE 700 1- Stantscheff R. Institute of Microbiology and Wine Research, Johannes Gutenberg University Mainz, Johann-Joachim Becherweg 15, 55128, Mainz, Germany aut NATIONALLICENCE 700 1- Müller L. Deutsches Biomasseforschungszentrum gemeinnützige GmbH (DBFZ), Torgauer Straße 116, 04347, Leipzig, Germany aut NATIONALLICENCE 700 1- Jacobi F. Deutsches Biomasseforschungszentrum gemeinnützige GmbH (DBFZ), Torgauer Straße 116, 04347, Leipzig, Germany aut NATIONALLICENCE 700 1- Polag D. Institute of Earthsciences, Ruprecht-Karls University Heidelberg, Im Neuenheimer Feld 234-236, 69120, Heidelberg, Germany aut NATIONALLICENCE 700 1- Keppler F. Institute of Earthsciences, Ruprecht-Karls University Heidelberg, Im Neuenheimer Feld 234-236, 69120, Heidelberg, Germany aut NATIONALLICENCE 700 1- König H. Institute of Microbiology and Wine Research, Johannes Gutenberg University Mainz, Johann-Joachim Becherweg 15, 55128, Mainz, Germany aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/16(2015-08-01), 6855-6863 0175-7598 99:16<6855 2015 99 253