This is a book review of 'Mycoplasma protocols: Methods in Molecular Biology, volume 104,' a comprehensive 33-chapter laboratory manual. Approximately half the volume covers practical laboratory techniques for mycoplasma isolation, propagation, and identification, with detailed media formulations and specimen treatment protocols—valuable for routine diagnostic laboratories. The second half addresses molecular and specialized techniques including immunodiagnostic methods, nucleic acid characterization (PCR, 16s rDNA sequencing, DNA-DNA hybridization), and advanced research methods such as transformation, transposon mutagenesis, and gene expression in mycoplasmas. The reviewer highlights chapters on metabolic studies and their relevance to recently sequenced Mycoplasma genomes, as well as protein characterization methods focusing on adhesins and surface antigens. Each chapter follows a consistent format with introduction, materials, methods, and practical tips from experienced authors. The book successfully provides technical detail to enable investigators to perform described protocols. While some specialized techniques like pulsed-field gel electrophoresis are omitted, the comprehensive coverage and practical emphasis make this a valuable laboratory reference for mycoplasma research and diagnosis.

Key findings

• The manual provides detailed protocols for mycoplasma isolation, identification, and characterization suitable for both routine diagnostic and specialized research laboratories
• Chapters on metabolic studies connect classical techniques to comparative genomic analysis using recently sequenced Mycoplasma genomes
• Specialized molecular methods covered include transformation, transposon mutagenesis, and novel gene expression approaches to overcome mycoplasma's atypical codon usage
• Practical tips and pitfalls based on author experience are included for each protocol to enhance usability