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Cultivation of microalgae provides an opportunity to solve the risk of fossil fuel utilization. The growth of microalgae is affected by various environmental factors such as light energy, pH, CO2, and nutrient supply. Among these factors, CO2 is the only carbon source for the growth of microalgae and serves for photosynthetic cell culture. Effectively delivery of CO2 is important for the culture of microalgae. Dunaliella tertiolecta is a marine green flagellate and fast growing alga. In this study focused on effect of carbon dioxide on microalgae, Dunaliella tertiolecta. Dunaliella tertiolecta UTEX LB 999 was cultivated at various CO2 concentrations and flow rates. The carbon dioxide concentrations were air, 0.3, 0.5, 1, 2, and 5%. The flow rates were 100, 200, 400, and 600cm³/min. The results indicated that CO2 concentrations more affected than flow rates for the growth of D. tertiolecta. The CO2 supplied to the photobioreactor affected the pH of the medium. The pH values were 7.80, 7.5, and 6.27, respectively, at air, 0.3, and 5% CO2 concentrations. Growth rates of D. tertiolecta increased with increasing concentration of CO2 and they are higher than without CO2 supplementation.