This detailed volume collects protocols from scientists who are actively engaged in developing or using protoplast technology. The book begins with chapters that focus on basic protoplast techniques and their utilities, such as protocols on protoplast isolation, transfection, and regeneration, as well as examples of how to use protoplasts for genome editing and gene function analysis in a number of major crop or model plant species. It continues by exploring protoplast automation, large scale functional genomics, and synthetic biology. 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.