Comparison of fiber samples from the 1994 Regional High Quality Tests (RHQT) were evaluated by conventional breeders' and Advanced Fiber Information Systems (AFIS) measurements. The fiber samples were from 18 current and potential varieties and were grown at seven locations in seven Southeastern states. Samples from two replications at each location were taken. The first objective was to estimate the proportion of the total fiber property variability that was due to genetic causes. The second objective was to compare the association of the AFIS and conventional breeder's fiber properties. All fiber properties showed significant genetic variability. Genetic variability as determined by AFIS for neps, short fiber content, maturity and fineness was readily detectable and offers the breeders a rapid method to determine these traits. Nep numbers were highly correlated with variety maturity, r = -0.75; fiber area, r = -0.70; and short fibers, r = 0.59. Fiber area and short fibers were also highly correlated, r = -0.65; and yarn tenacity from conventional breeders' evaluations was highly correlated with fiber area, r = -0.76. Of greatest concern to breeders was the association of fiber area with neps and short fiber content. Unanswered but important questions that need to be addressed in the future are the genetic association of AFIS fiber properties with spinning efficiency, quality, and agronomic acceptability.