In this video, we present the procedures used for making crosses in Upland Cotton, Gossypium hirsutum. These are methods performed by the Molecular Cotton Breeding Lab in Tifton, GA. Making crosses between cotton lines is a very important step within a breeding program to improve yield, fiber quality traits, and disease resistance. This form of crossing cotton plants can be beneficial to a wide range of breeders in a variety of programs to ensure efficient and pure crosses of their lines.