This collection of experimental and computational strategies and techniques for microbial genome-scale essentiality studies was developed and written by the leading research groups in the field. In addition to wet-lab protocols, the book describes statistical methods essential for planning and evaluating successful large-scale essentiality screens; in-silico prediction of gene essentiality using genome-scale reconstructed metabolic models; and data integration and comparative analysis of genomic databases. Each protocol follows the successful Methods in Molecular Biology(TM) series format, offering an introduction outlining the underlying principles, step-by-step instructions, a list of necessary equipment and reagents, and tips on troubleshooting and avoiding pitfalls.