The composition, variation, and availability of feedstocks for biofuel production vary significantly across different geographical regions, impacting both the feasibility and sustainability of biofuel technologies. This chapter explores diverse feedstock options utilized in biofuel production, including first-generation, second-generation, and third-generation sources, and examines how their availability is shaped by regional agricultural practices, environmental factors, and socioeconomic conditions. First-generation biofuels, derived from edible crops such as sugar, starch, and oils, have been widely used but face challenges relating to food security, land use competition, and environmental degradation. In contrast, second-generation and third-generation biofuels, sourced from nonfood feedstocks like algae, agricultural residues, and municipal waste, present a more sustainable alternative but encounter hurdles due to feedstock complexity and processing challenges. The regional variation in feedstock composition—driven by climate, land availability, and local agricultural systems—plays a crucial role in determining the success and scalability of biofuel production systems. Furthermore, developing advanced biotechnological processes is essential for enhancing the efficiency of feedstock conversion and improving the overall sustainability of biofuel production. Policy frameworks tailored to regional contexts are also vital in promoting the adoption of sustainable feedstocks and encouraging innovation in biofuel technologies. This chapter provides a comprehensive analysis of the global landscape of feedstock availability for biofuels, highlighting the challenges and opportunities inherent in regional differences and offering insights into the pathways for optimizing biofuel production systems worldwide.