In addition to high yields, improving the quality of cotton (Gossypium hirsutum L.) fiber has become an increasingly important component of the value of cotton, especially for marketing in the international trade. The present study was designed to identify genetic sources of variation in F₂ populations for improving cotton fiber quality traits. This material will be genotyped with microsatellite markers, and the genotypic and phenotypic information combined to detect quantitative trait loci. Seven Upland cotton cultivars (TM1, 7235, SG125, Fibermax832, CAMD-E, MD51, and DPL50) were selected to represent a range in fiber quality and productivity. They were paired in a diallel design to obtain 21 F₁ crosses, which were used to develop F₂ populations. In 2004, 200 plants from each of the 21 F₂ populations along with their respective parents were measured to obtain yield and fiber data. In addition, leaf tissue was collected from individual F₂ and parent plants for molecular marker analysis. Results of the phenotypic data show that F₂ populations significantly differ for 2.5% span length (P = 0.0184) with (TM1 x 7235)F₂ having the greatest length (1.20mm) and (Fibermax832 x CAMD-E)F₂ and (CAMD-E x DPL50)F₂ having the shortest length (1.05mm). Micronaire also differed among the F₂ populations (P = 0.0215). (7235 x SG125)F₂ and (7235 x Fibermax832)F₂ had the lowest micronaire (3.8 units) while (SG125 x Fibermax832)F₂ had the highest micronaire (5.2 units). There was a slight difference among F₂ populations for elongation (P = 0.098). The smallest elongation was measured in (TM1 x Fibermax832)F₂ (6.0%) and (CAMD-E x DPL50)F₂ (6.0%).  (MD51 x DPL50)F₂ and (7235 x MD51)F₂ had the greatest elongation (8.1%). These preliminary results indicate that there is variation for several cotton fiber measurements among these F₂ populations that will be useful in identifying regions of the cotton genome specific for fiber quality traits.