Veggie NEFF media for Tetrahymena thermophila v1

The ciliate Tetrahymena thermophila has been a research organism used by a great community of scientists including biologist, geneticists and toxicologists. T. thermophila is a bacterial predator; however, we can grow it in the lab without bacteria, in axenic media, if needed. The media presented here is a modification of the axenic NEF media published in the Tetrahymena stock center (RRID:SCR_008362, https://tetrahymena.vet.cornell.edu/recipes.php) under modified NEF media, and originally published by Cassidy-Hanley D, Bowen J, Lee JH, Cole E, VerPlank LA, Gaertig J, Gorovsky MA, Bruns PJ. Germline and somatic transformation of mating Tetrahymena thermophila by particle bombardment. Genetics. 1997 May;146(1):135-47. doi: 10.1093/genetics/146.1.135. PMID: 9136007; PMCID: PMC1207932. We have modified the media to use peptone free from animal products. The original component "Proteose Peptone" is the processed result of animal products, which brings high variability associated with each production lot every time we buy a new product. Because of this, we have been growing cells and testing some behaviors while using peptone from soy. This is the media we present here. The protocol has three sections: First part describes how to prepare NEFv (NEF media with "vegetarian" peptone from soy and without iron complement), the second describes how to prepare the stock concentration of iron chloride and its storage, and the third describes how we get all components together just before we use the media.