This comparative study investigates the production of biofuel from the pyrolysis of corn stover, where three different case studies were analyzed and compared to explore the potential of utilizing and optimizing the resulting biofuels as an alternative energy source. The effect of temperature on the product distribution and the biofuel properties was a major objective in this paper. Moreover, different reactors of different heating methods were evaluated along with other main variables such as particle sizes, the presence of catalysts as bed materials, acid and steam explosion pretreatments were also explored. The optimum temperature ranges for maximum bio-oil production are found to be 400 °C and 500 °C. Also, the highest bio-oil yield and HHV were obtained at 400 °C, which were about 43.3 wt% and 33.8 MJ/kg, respectively, from fast pyrolysis of corn stover in a catalytic fluidized bed reactor. While the highest biogas yield was about 73 wt% which was achieved by the microwave pyrolysis of corn stover, this reactor design also improved biogas properties but produced low bio-oil and biochar yields. Acid pretreatment had a positive effect on bio-oil yield using microwave pyrolysis, however reducing the particle size had an opposite effect. The products and feedstock properties such as pH level, ash content, volatile matter, and moisture were all analyzed by different standard analyses such as the GC–MS analysis and FTIR analysis. The elemental analysis was also performed to find the most significant compounds in feedstock and products in each case study.