Energy use increasing is caused by increasing population and lack of reserves to produce oil, and also problem of emissions from fossil fuels will put pressure on every country to be able to produce renewable energy. Biogas is a renewable energy that can be solution to alternative energy needs. This research aims to determine the effect of glycerol utilization in making biogas from cow dung. This research method used experimental method. The results showed that 0% glycerol temperature was 28.56°C, 5% glycerol temperature was 29.7°C, 10% glycerol temperature was 30.16°C and 15% glycerol temperature was 30.5°C, with upper pH of 0% glycerol was the lowest pH of 5.389 and 15% glycerol was the highest pH of 6.61, while lower pH of 0% glycerol was the lowest pH of 5.49 and also the highest pH was found in 15% glycerol which was 6.58. At 0% glycerol biogas volume yields 18.53 ml, 5% glycerol yields 25.18 ml, 10% glycerol yields 31.89 ml, and at 15% glycerol yields 55.51 ml. In the 0% glycerol flame test, it produces 46 seconds, 5% glycerol produces 1 minute 15 seconds, 10% glycerol produces 2 minutes 48 seconds and 15% glycerol produces 3 minutes 8 seconds.