Microalgae Phaeodactylum tricornutum was grown in different conditions to increase lipid content for biodiesel production. Adding a carbon source like glycerol boosted lipid production, with the best results seen when glycerol was added at the start. Salinity of 15/00 promoted the highest algae growth. Glycerol at 0.1 M concentration led to cultures with 1.3 g/L of biomass. Mixotrophic cultivation with 24 hours of light and glycerol resulted in 80% more lipids compared to autotrophic growth.