The genotypes evaluated in this study was a part of the National Screening Program for Fusarium Wilt Resistance. This program is conducted annually in the greenhouses of Cotton and Fiber Crops Diseases Research Dept., PPRI. The present study included 50 of Upper Egypt experimental genotypes and 875 families of six commercial cultivars. The experimental genotypes were supplied by Cotton Breeding Dept., and the six commercial cultivars were provided by Cotton Maintenance Dept., Cotton Research Institute. Healthy survival rate of seedlings of the genotypes was used as a criterion to evaluate the reactions when the genotypes were grown in autoclaved highly infested soil with a mixture of race 3 of Fusarium oxysporum f.sp.vasinfectum isolates. The majority of the tested genotypes (48%) were very highly resistant while 22% were very highly susceptible. All the families of the commercial cultivars Giza 90, Giza 95, Giza 86, Giza 87, and Giza 88 were very highly resistant while only 99.30% of Giza 92 families showed this level of resistance, which deserves more attention in monitoring its families in the future to Avoid any varietal deterioration. The present work provides new and useful sources of resistance to FOV race 3 isolates.