Mycobacteria Protocols, Second Edition is a comprehensive reference book that focuses on the biology of mycobacteria, particularly in light of the growing threat posed by tuberculosis and other mycobacterial infections. This updated edition refines the methods from the first edition and integrates newly developed techniques derived from the latest research in the field. The chapters cover a wide range of topics, from basic procedures for subcellular fractionation to advanced methods that include specific growth conditions as well as analyses of the whole genome, transcriptome, and proteome. The structure follows the established Methods in Molecular Biology series and provides detailed laboratory protocols, material lists, and helpful troubleshooting tips. This book is a valuable resource for researchers looking to advance the understanding and treatment of mycobacteria.