Laminar burning speed and ignition delay time behavior of iso-octane at the presence of two different biofuels, ethanol and 2,5 dimethyl furan (DMF), was studied in this work. Biofuels are considered as a better alternative source of fossil fuels. There is a potentiality that combustion characteristics of iso-octane can be improved using biofuels as an oxygenated additive. In this study, three different blending ratios of 5%, 25%, and 50% of ethanol/iso-octane and DMF/iso-octane were investigated. For laminar burning speed calculation, equivalence ratio of 0.6–1.4 was considered. Ignition delay time was measured under temperature ranges from 650 K to 1100 K. Two different mechanisms were considered in numerical calculation. These mechanisms were validated by comparing the results of pure fuels with wide range of experimental and numerical data. The characteristic change of iso-octane with the presence of additives was observed by comparing the results with pure fuel. Significant change was observed on behavior of iso-octane at 50% blending ratio. A comparison was also done on the effect of two different additives. It has found that addition of DMF brings significant changes on iso-octane characteristics comparing to ethanol.