Cotton is a crop with significant global impact that produces fiber, oil and protein. Cotton is a multi-purpose crop that supplies three primary products viz., cotton lint, linters and cotton seed. For development of new Bt hybrids first we would need to identify stable non-Bt hybrids and then convert the best hybrid parents into Bt version. The present investigation entitled "Genetic analysis in cotton (Gossypium hirsutum L.)" was undertaken to generate information on the magnitude of heterobeltiosis and standard heterosis, gene effects, combining ability effect of parents and hybrids for seed cotton yield and its component characters and genetic diversity of parental genotypes using SSR markers. The experimental material comprised of sixteen diverse females and males of G. hirsutum and their resultant 64 F1s developed through Line × Tester mating design along with one standard check (G. Cot. Hy. 18) were evaluated in randomized complete block design with three replications at Regional Research Station, Anand Agricultural University, Anand during Kharif-2022.