To improve the accuracy of the discrete element research, physical and simulation experiments were used to calibrate the cotton stalk contact parameters. Based on the stalk-stalk and stalk-steel contact mechanics, the parameters were measured in physical experiments, and the discrete element simulation software was used to build the stalk model. In the simulation process, the Plackett-Burman experiment was used to screen three significant factors from six initial factors. The steepest Plackett-Burman experiment was used to determine the optimal interval of the significant factors. A second-order regression model of the significant factors and the angle of repose was established according to the Central Composite design experiment. The best parameter combination of the significant factors was then obtained: the coefficient of static friction on stalk-steel contact was 0.31, the coefficient of static friction on stalk-stalk contact was 0.62, and the coefficient of rolling friction on stalk-stalk contact was 0.02. The relative error between the physical angle of repose and the simulated angle was 3.27%, indicating that it is feasible to apply the simulation experiment instead of the physical one. It offers insights into cotton stalk contact parameter settings and film-stalk separation in the simulation.