This book explores cutting-edge methods to work with the notoriously difficult, but highly prevalent, obligate intracellular pathogen, Chlamydia trachomatis. These include techniques to identify Chlamydia trachomatis in patient samples, ranging from simple point-of-care tests to whole genome sequencing; methods for propagation of strains in both cell culture and animal models; techniques to manipulate Chlamydia trachomatis in molecular genetic methodologies; a high-throughput screening method for testing new potential drugs against intracellular bacteria; a screen for antibiotic resistance; methods for labeling and enumeration; and descriptions of genotyping technologies, as well as dual RNA-Seq transcriptional profiling. 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.