The USDA-ARS Pee Dee germplasm enhancement program was initiated in 1935. Although initially focused on developing Sea Island cultivars for the southeast US, the program's long-term objective has been to develop high yielding Upland germplasm with Sea Island fiber properties. The foundation of the Pee Dee program was developed using an array of unique breeding methods and diverse germplasm that included Sea Island, Upland, and triple hybrid strains. Since 1935, the Pee Dee program has released greater than 80 cultivars and germplasm lines. Beginning in 2004, a series of experiments were initiated to examine genetic properties of the Pee Dee germplasm collection. These experiments examined a number of genetic properties of the Pee Dee germplasm program including: 1) molecular and phenotypic trait variation, 2) genetic gain, 3) correlated traits, 4) genotype-by-environment interactions, and 5) breeding potential. Results of these experiments will be discussed with a focus toward future breeding applications.