This second edition provides new and updated chapters describing the utility and attributes of different tools that are used for studying microbial systems. Chapters detail a number of methods, including in silico system level analyses by MetaFlux, Kbase genome-scale model builder, COBRA toolbox, NanoSIP, and PAMMS. Written in the highly successful series format, 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, aims to introduce and aid scientists in using the various tools that are currently available for analysis, modification, and utilization of microbial organisms.