The study presents a detailed protocol for reproducibly reconstructing eleven major white matter tracts in the human brain using diffusion tensor imaging (DTI). The protocols were developed through iterative refinement of intra- and inter-rater measurements and were tested by raters without prior tractography experience. The protocols were applied to a database of normal adult subjects to assess tract size, fractional anisotropy (FA), and T2 values. Distinctive features in FA and T2 were found for the corticospinal tract and callosal fibers, while hemispheric asymmetry was observed in white matter tracts projecting to the temporal lobe. The study demonstrates that the established protocols provide reliable and reproducible DTI-based tract-specific quantification. The reproducibility of the protocols was evaluated using κ statistics, which showed high agreement between raters. The results indicate that the protocols are robust and can be used for quantitative analysis of white matter tracts. The study also highlights the importance of protocol setup and measurement of reproducibility in ensuring the accuracy and reliability of tractography results. The findings suggest that tractography can be a valuable tool for studying white matter anatomy and its changes in disease. The study provides guidelines for reproducible DTI-based tract-specific quantification and emphasizes the need for further research to improve the accuracy and reliability of tractography methods.The study presents a detailed protocol for reproducibly reconstructing eleven major white matter tracts in the human brain using diffusion tensor imaging (DTI). The protocols were developed through iterative refinement of intra- and inter-rater measurements and were tested by raters without prior tractography experience. The protocols were applied to a database of normal adult subjects to assess tract size, fractional anisotropy (FA), and T2 values. Distinctive features in FA and T2 were found for the corticospinal tract and callosal fibers, while hemispheric asymmetry was observed in white matter tracts projecting to the temporal lobe. The study demonstrates that the established protocols provide reliable and reproducible DTI-based tract-specific quantification. The reproducibility of the protocols was evaluated using κ statistics, which showed high agreement between raters. The results indicate that the protocols are robust and can be used for quantitative analysis of white matter tracts. The study also highlights the importance of protocol setup and measurement of reproducibility in ensuring the accuracy and reliability of tractography results. The findings suggest that tractography can be a valuable tool for studying white matter anatomy and its changes in disease. The study provides guidelines for reproducible DTI-based tract-specific quantification and emphasizes the need for further research to improve the accuracy and reliability of tractography methods.