The increasing global demand for biofuels for energy security and to reduce the effects of climate change has created an opportunity to explore new sources of biomass, of which, microalgae is the most promising one. The Laboratory of the Biomass Research Centre (CRB, University of Perugia) is equipped with a photobioreactor that is used to cultivate microalgae under batch conditions. Tests were carried out a temperature of 22 °C and a Photosynthetic Photon Flux Density of 140 µE·m−2·s−1. Cultures were characterized in terms of biomass produced and lipid fraction distribution. The novelty of this paper is the measure of the fuel properties of Selenastrum capricornutum, a new strain for biodiesel production. In particular, after the microalgae have been collected and oil has been extracted, this has been transesterified using a methanol/NaOH solution. The resulting biodiesel has been analyzed with a high-resolution gas chromatograph to determine the concentration of the different methylesters.