Cyclotrons are currently used for the preparation of a wide variety of radionuclides that have applications in single photon emission computed tomography (SPECT) and positron emission tomography (PET). Consequently, there is high demand from IAEA Member States for support in the area of radiopharmaceutical production using cyclotron produced radioisotopes. This publication describes the potential radionuclide production routes using cyclotrons in different energy ranges and provides methods for the development of targets and provides details of the chemistry for the separation of radionuclides from target materials. The readership of this publication includes scientists, operators interested in putting this technology into practice, technologists already working with cyclotrons who wish to enhance the utility of existing machines, and managers in the process of setting up radionuclide facilities in their countries. Students working towards higher level degrees in related fields may also benefit from this publication.