This chapter describes a stepwise protocol for Agrobacterium -mediated sorghum genetic transformation. Immature embryos from sorghum plants were used as the target explants. The Agrobacterium strain LBA4404, carrying a “super-binary” vector, was used in this protocol. Agrobacterium co-transformation vectors, one T-DNA containing the selectable marker gene and another T-DNA containing the trait gene(s), were also introduced in sorghum transformation for eliminating the selectable marker gene in the resulting transgenic plants. This chapter provides recommendations for analysis of the transgenic plants to confirm T-DNA integration into the sorghum genome and segregation of the selectable marker gene from the trait gene(s).