Resumo. The objective of this work was to set the adequate amount of sample matter, the substrate type and the ideal temperature to perform the germination test of Eucalyptus dunnii in laboratory. Seeds, from Telêmaco Borba, PR, were purchased at Klabin Paraná Papéis. Two experiments were performed in the Seed Improvement Laboratory of the Pontific Catholic University of Parana (PUCPR). The first experiment was performed to determine the ideal amount of sample matter for the germination test. It had five treatments and four replications. The five treatments were different sample matters (365, 730, 1.095, 1.460 and 1.825 mg) from the lot, which corresponded to 50, 100, 150, 200 and 250 mg of pure seeds. The number of germinated seeds per gram of the samples was evaluated after 14 days. In the second experiment, the influence of paper, sand and vermiculite substrates and temperature (20, 25 or 30ºC) on seed germination was verified. Percent germination (%), average germination time (days), average germination speed (day-1), index of germination speed and entropy (bits) were analyzed. It was concluded that seed germination of Eucalyptus dunnii may be performed with sample matter equivalent to 50 mg of pure seeds. Sand or paper must be used for the germination test. A better performance of the Eucalyptus dunni seed population was observed at 20 ºC.