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This detailed book provides practical information for the laboratory that can be applied to the study of vaccinia and other poxviruses while emphasizing long-standing field standards and focusing on emerging new technologies applied in the field of poxvirology. The methods and protocols have been designed with the bench scientist in mind, being presented in a fashion that makes them useful for both starting and veteran poxvirus researchers. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Vaccinia Virus: Methods and Protocols serves as a valuable resource for scientists looking to bring new methods and procedures into their lab in order to make exciting discoveries that will continue to deepen our understanding of this fascinating virus family.
List of contents
Working Safely with Vaccinia Virus: Laboratory Technique and Review of Published Cases of Accidental Laboratory Infections with Poxviruses.- Bioinformatics for Analysis of Poxvirus Genomes.- Simple, Rapid Preparation of Poxvirus DNA for PCR Cloning and Analysis.- Construction and isolation of recombinant vaccinia virus expressing fluorescent proteins.- Generation of Vaccinia Virus Gene Deletion Mutants Using Complementing Cell Lines.- Vaccinia genome editing using CRISPR.-RNAi-mediated depletion of poxvirus proteins.- Assessing the Structure and Function of Vaccinia Virus Gene Products by Transient Complementation.- Preliminary Screening and In vitro confirmation of orthopoxvirus antivirals.- Vaccinia virus transcriptome analysis by RNA sequencing.- Ribosome profiling of vaccinia virus-infected cells.- Quantitative PCR-based Assessment of Vaccinia Virus RNA and DNA in Infected Cells.- Click Chemistry Based Labelling of Poxvirus Genomes.- Visualizing poxvirus replication and recombination using live cell imaging.- High-Content Analyses of Vaccinia Plaque Formation.- Super-Resolution Microscopy of Vaccinia Virus Particles.- Bioluminescence imaging as a tool for poxvirus biology.- Growth and Purification of Vaccinia Virus Stocks for Intravital Imaging.- Intravital Imaging of Vaccinia Virus-Infected Mice.
