For the biodiesel fuel production from microalgae, the lipid from wet and dry samples of green algae Scenedesmus sp. was extracted by using various solvents and pre-treatment methods. Extraction yield of the lyophilized sample was better than that of dry sample. Chloroform/methanol (2:1, v/v) and ultrasonication or homogenization method were also selected as the most effective solvent and pre-treatment methods for lipid extraction, respectively. Under these constraint conditions, optimization experiment of lipid extraction was investigated by Taguchi approach using orthogonal matrix $L_9$ ($3^4$) method. The optimum extraction conditions of lipid extraction was obtained at pre-treatment of homogenization, extraction time of 5 hour, temperature of $35^{circ}C$, and solvent ratio of 1:20 (w/v). Yield of extraction at optimized condition was 20.55% and it was 96% of total lipid content (21.38%) of Scenedesmus sp.