For the last two decades, cotton breeders have used High Volume Instrument (HVI) as their primary and often sole source of fiber quality data when making plant selections. Fiber data generated by Advanced Fiber Information System (AFIS) technology is also now available to plant breeders, and provides additional information on length characteristics and fiber maturity. Two methods of evaluating fiber quality of upland cotton (Gossypium hirsutum L.) in a breeding program were compared. One method used only HVI data for plant selections, whereas the other method used only AFIS data. One critical difference between the selection methods was use of fiber length distributions as the only selection criteria in the AFIS method. Line development began with 15 intraspecific F2 populations in 2005. Selections were made in the F2 and F3 generations based upon data generated from either HVI or AFIS. In the F5 generation,10 lines from each selection method and four commercial cultivars were planted at three locations. Improvement of fiber quality in selected lines was apparent in the F3 generation. Both selection methods resulted in F5 lines with better fiber quality than commercial cultivars. Fiber maturity had a significant impact on length characteristics. Data indicated it is possible to improve fiber length distribution using either selection method. Principle component analysis revealed differences between length distribution of HVI selection lines and AFIS selection lines, even though average fiber properties from each selection method were similar.