This is the author manuscript accepted for publication and has undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record. Please cite this article as doi: 10.1002/cpmc.110. This article is protected by copyright. All rights reserved. [*This is a protocol for inclusion in CPMC volume 58. There are 2 figures and 1 table included. There is also a required PMC file included.] Article Title: The Propagation, Quantification, and Storage of Vesicular Stomatitis (VSV) AUTHOR(S) AND CONTACT INFORMATION: Alaa A. Abdelmageed (aaa9470@g.rit.edu) and Maureen C. Ferran (mcfsbi@rit.edu)* Address: Thomas H. Gosnell School of Life Sciences, Rochester Institute of Technology, Rochester, NY 14623 *Corresponding author: Thomas H Gosnell School of Life Sciences, Rochester Institute of Technology, 153 Lomb Memorial Drive, Rochester, NY 14623, USA. E-mail address: mcfsbi@rit.edu (M.C. Ferran) ABSTRACT Vesicular stomatitis virus (VSV) is the prototypical member of the Rhabdoviridae family of negative sense single-stranded RNA viruses. This virus has been used as a powerful model system for decades and is currently being used as a vaccine platform and an oncolytic agent. Here we present methods to propagate, quantitate, and store VSV. We also review the proper safety protocol for the handling of VSV, which is classified as a Biosafety Level 2 pathogen by the United States Centers for Disease Control and Prevention. Basic Protocol 1: (GENERATION, PURIFICATION, AND STORAGE OF VSV VIRUS STOCKS) Basic Protocol 2: (QUANTIFICATION OF VSV BY PLAQUE ASSAY) Support Protocol 1: PROPAGATION OF VERO CELLS KEYWORDS: Vesicular stomatitis virus, stock, propagation, titration, plaque assay, storage. INTRODUCTION Vesicular stomatitis virus (VSV) is an enveloped, non-segmented, negative-sense RNA virus belonging to the Rhabdoviridae family. Its genome (~11-kB in length) encodes five proteins: the surface glycoprotein (G), the matrix protein (M), the nucleocapsid protein (N), the phosphoprotein (P), and the RNA-dependent RNA polymerase (L) (Wagner & Rose, 1996).