AbstractThis study evaluates uncertainties in the techno‐economic analysis of transportation fuel production from biomass gasification and mixed alcohol synthesis. Two scenarios are considered: a state‐of‐technology scenario and a target scenario with projected technological advances. Uncertainties of more than 10 parameters are investigated. The probability distributions of these parameters are estimated based on historical price data and experimental data. Data samples generated from the corresponding distribution are then utilized to run a Monte Carlo simulation. The results yield minimum fuel‐selling prices of $ 1.85 L−1 with a standard deviation of 0.13 for the state‐of‐technology scenario and $ 1.14 L−1 with a standard deviation of 0.11 for the target scenario, respectively. The feedstock price and internal rate of return (IRR) have significant impacts on the minimum fuel‐selling price in both scenarios. These findings are indicative of the reduction in biofuel cost and uncertainty achievable with increasing technology maturity.