The most relevant way for phytoplasma transmission, both in nature and in the laboratory, is via insect-mediated inoculation. Therefore, the setup of proper transmission trials is necessary to investigate vector–phytoplasma interactions under controlled conditions. Here we present protocols for laboratory transmission experiments with insect vectors, describing procedures for measuring the acquisition, latency, and inoculation phases. The use of plant sources from the field is presented along with the use of experimentally infected laboratory plants. Similarly, methods to measure the field infectivity of insects are discussed in conjunction with experiments involving laboratory-reared specimens for studies on all transmission phases. Inoculation is described involving the use of an artificial diet, the final host plant, or a model plant.