Protocol

Mass Culture of Paramecium tetraurelia

¹ Centre de Génétique Moléculaire, Centre National de la Recherche Scientifique, FRE3144, F-91198 Gif-sur-Yvette, France
² Laboratoire de Biologie Cellulaire 4, Centre National de la Recherche Scientifique, UMR 8080, Université Paris-Sud, 91405 Orsay Cedex, France
³ Laboratoire de Génétique Moléculaire, Centre National de la Recherche Scientifique, UMR 8541, École Normale Supérieure, F-75230 Paris, France
⁴ Laboratoire de Biométrie et Biologie Évolutive, Centre National de la Recherche Scientifique, UMR 5558, Université Lyon 1, F-69622, Villeurbanne, France
⁵ Laboratory of Molecular Biology and Department of Genetics, University of Wisconsin-Madison, WI 53706, USA
⁶ Department of Biology, Indiana University, Bloomington, IN 47405-3700, USA

⁷Corresponding author (emeyer{at}biologie.ens.fr).

INTRODUCTION

Clonal cell lines of Paramecium tetraurelia can be established easily by single cell isolation. Given a constant supply of food, P. tetraurelia cells will remain in the vegetative phase of their life cycle and, at 27°C, will divide by binary fission every 6 h. P. tetraurelia is suitable for a range of biochemical and molecular studies such as mitochondrial genetics, post-translational tubulin modifications, variant genetic codes, programmed genome rearrangements (and their epigenetic regulation by noncoding RNAs), and RNA interference. This protocol describes the methods required to grow high-density mass cultures of P. tetraurelia in quantities sufficient to provide the material required for biochemical and molecular biological studies.