In this study, the harvesting of a biomass from a high rate algal pond (HRAP) of a real-scale domestic wastewater treatment facility and its potential as a biomaterial for the production of biodiesel were investigated. Increasing the medium pH to 12 induced high flocculation efficiency of up to 96% of the biomass through both sweep flocculation and charge neutralization. Lipids extracted by ultrasounds from this biomass contained around 70% of fatty acids, with palmitic and stearic acids being the most abundant. The extract obtained by supercritical CO2 contained 86% of fatty acids. Both conventional solvents extracts contained only around 10% of unsaturated fats, whereas supercritical CO2 extract contained more than 40% of unsaturated fatty acids. This same biomass was also subject to direct extractive-transesterification in a microwave reactor to produce fatty acid methyl ester, also known as, raw biodiesel.