Positron emission tomography (PET) with 68Ga is a fast-growing field in molecular imaging, both in research and in clinical routine. The availability of 68Ga via the 68Ge/68Ga radionuclide generator facilitates the development and production of radiopharmaceuticals independent of a cyclotron. The presented work shows a complete 68Ga labelling study exemplified on [68Ga]DO2A-(butyl- l-tyrosine)2, a potential tumour tracer for PET. A methodical sequence is followed to optimize the 68Ga-labelling reaction. Practical aspects are described and the different parameters contributing to the labelling yield are demonstrated. The influence of temperature, time, amount of labelling precursor and pH value on the radiochemical yields is demonstrated. A conventional heating method is compared with microwave irradiation as an alternative labelling method. Finally, purification of 68Ga-labelled compounds via solid-phase extraction and quality control is shown. The procedure described in this manuscript may serve as a guideline for optimizing 68Ga labelling reactions. Copyright © 2011 John Wiley & Sons, Ltd.