There are several media and supplements are available in the market for growing microalgae which are artificial, and made for growing microbes. Cow urine is a natural nutritional source for growing algae. It observed while visiting cow farms in the village and for getting surety work has carried out. Microalgae for biofuel is a topic of recent interest as it has overcome the problems related to the previous two generations of biofuel, i.e. freshwater and fertile land availability which was driving food prices high. Selected microalgae species has studied for ten days consistently for analysing its growth with cow urine. And it has observed that a 10% cow urine supplemented culture shows better biomass yield 2.134 g/L while for BG11 broth it is 0.734 g/L.