Supplementary Material New insights into the ecology and physiology of Methanomassiliicoccales from terrestrial and aquatic environments. Marc Cozannet1, Guillaume Borrel2, Erwan Roussel1, Yann Moalic1, Maxime Allioux1, Amandine Sanvoisin1, Laurent Toffin1 & Karine Alain1,* 1 Univ Brest, CNRS, IFREMER, IRP 1211 MicrobSea, Laboratoire de Microbiologie des Environnements Extrêmes LM2E, UMR 6197, IUEM, Rue Dumont d’Urville, F-29280 Plouzané, France;
[email protected] (M.C.);
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[email protected] (K.A.) 2 Department of Microbiology, Unit Evolutionary Biology of the Microbial Cell, Institut Pasteur, Paris, France;
[email protected] (G.B.) * Correspondence:
[email protected]; Tel.: +33-0298-4988-53 Received: date; Accepted: date; Published: date Supplementary Text Text S1. Pre-metabarcoding screening Section 1.1. Total DNA extraction In order to search for natural samples containing Methanomassiliicoccales, an initial large-scale molecular screening was carried out on a wide range of anoxic samples (86 in total). DNA extractions were carried out in triplicate with a protocol adapted to each type of matrix. Two types of DNA extraction procedures have been implemented depending on the nature of the matrices: (i) Nucleic acids were extracted from sediments, mud and peatland soils by combining a mechanical lysis step to a commercial DNA extraction kit for soil type matrices. The first extraction step consisted of mechanical cell lysis of 0.5 g of sample by bead-beating (5500 rpm, 30 s), using the Precellys 24® tissue homogenizer (Bertin instruments, France).