Background/Objectives: The demand of energy isincreasing exponentially worldwide. This demand is mostly supplied by the fossil fuels, which is a nonrenewable source of energy. The most recently, renewable source of energy, Microbial Fuel Cell (MFC) is attracting the scientist over the globe for research. Methods/Statistical Analysis: MFC is a promising green technology which simultaneously generates direct electricity through metabolic activities of the microbes and also treating the organic waste. This technology is not commercialized yet, but holds a promising future, both in terms of energy production and waste reduction. In the present work, a Single chamber single electrode Microbial Fuel Cell (MFC) has been fabricated to generate electricity from sewage sludge, cow dung and kitchen waste at an ambient temperature of 32±10°C. The performance on the basis of voltage obtained, surface power density and COD reduction have been evaluate for those different substrate on same ambient condition. Findings: The change in pH is also recorded for all the substrate used. The maximum voltage output of 1652 mV is obtained with sewage sludge on the 5th day (120 hrs.) with the surface power density of 988.32 mW/m2. The maximum voltage obtained from kitchen waste is about 657 mV on 8th day (192hrs.) and in case of cow-dung the maximum voltage is about 452 mV on 8th day (216hrs.). Almost constant output voltage of about 350 mV for a month can be harvested with cow dung. At the end of the 29th day, the maximum COD reduction for the cow dung was 50%. Applications/Improvements: When comparing with other single chamber single electrode MFC, the present model is generating more electricity that any MFC using sewage sludge as substrate except platinum electrode, which is much costlier that electrode used in the present study.