This study describes protocols for reconstructing eleven major white matter tracts using diffusion tensor imaging (DTI). The protocols were refined through iterative intra- and inter-rater measurements and identification of sources of variability. Reproducibility of the established protocols was tested by raters with no prior experience in tractography. The protocols were applied to a DTI database of adult normal subjects to study the size, fractional anisotropy (FA), and T2 of individual white matter tracts. Distinctive features in FA and T2 were observed for the corticospinal tract and callosal fibers. Hemispheric asymmetry was noted for tracts projecting to the temporal lobe. The study provides guidelines for reproducible DTI-based tract-specific quantification.This study describes protocols for reconstructing eleven major white matter tracts using diffusion tensor imaging (DTI). The protocols were refined through iterative intra- and inter-rater measurements and identification of sources of variability. Reproducibility of the established protocols was tested by raters with no prior experience in tractography. The protocols were applied to a DTI database of adult normal subjects to study the size, fractional anisotropy (FA), and T2 of individual white matter tracts. Distinctive features in FA and T2 were observed for the corticospinal tract and callosal fibers. Hemispheric asymmetry was noted for tracts projecting to the temporal lobe. The study provides guidelines for reproducible DTI-based tract-specific quantification.