Introduction. This protocol describes a method for obtaining in vitro germination of zygotic embryos from open-pollinated wild seedy bananas and controlled hybridizations. The principle, key advantages, starting plant material, time required and expected results are presented. Materials and methods. This part describes the required laboratory materials, medium preparation, embryo extraction and culture. Results. Embryos typically begin germination in the dark about 15 days after inoculation on germinating medium. After 30––40 days of culture, plantlets are subcultured on a plant growth medium and placed in the light to achieve plant growth and rooting. The ratios of zygotic embryos obtained vary with the types of crosses achieved between Musa sp. genotypes.