Recently, algae seem to be a promising feedstock for the future alternative fuel (green fuel) instead of fossil fuel. Algae (macro- and micro-algae) may represent as a main resource for biofuel and biodiesel due to their high biosynthesis effeminacy to produce lipids and fatty acids comparing to other types of biomass. Algae can be an interesting source of commercial and eco-friendly fuel due to algae local availability and low CO₂ emission, respectively. The current study was conducted to compare the percentage of oil production in three different genera of algae, including Cosmarium, Spirogyra and Chara, and examine the ability of using the algae oil as an alternative fuel for industrial purposes. Soxhlet-based method and n- hexane organic solvent were used for extraction of algae oils followed by biochemical analysis and gas chromatography (GC-Mass) technique for algae oil detection. The algae Cosmarium had 2.8 % algal oil yield,  and recorded the highest yield of the total fatty acids (65.60 %), whereas Spirogyra recorded a low result by producing  32.07 %, noting that Chara alga showed the lowest value (21.58). The chemical analysis of algal oils showed that the quality of algae oil fatty acids proved to be the best choice. Cosmarium oil recorded the highest ratio (2.05 %) for unsaturated fatty acids (USAFA)/ saturated fatty acids (SAFA) and the degree of unsaturation (DU) was 84.86. While, Spirogyra and Chara showed that the value of USAFA/SAFA was 1.58 and 0.56 % with DU of 27.74 and 9.68, respectively. In this context, Cosmarium alga proved to be the best feedstock for biomass of algal oil production with astounding properties of algal biodiesel.