This detailed volume presents experimental and computational methods for constructing and interrogating synthetic gene circuits across multiple organizational scales. Beginning with a section on genetic tools, platforms, and experimental frameworks that enable sophisticated circuit construction, the book continues by covering methods for regulating circuit behavior and mitigating variability, as well as frameworks for understanding time-dependent circuit behavior in growing cells and populations. 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 and readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls. Authoritative and practical, Synthetic Gene Circuits: Methods and Protocols aims to support a synthetic biology practice that is predictive by design, robust by control, and interpretable through dynamics.