Connectional neuroanatomical maps can be generated in vivo by using diffusion-weighted magnetic resonance imaging (dMRI) data, and their representation as structural connectome (SC) atlases adopts network-based brain analysis methods. We explain the generation of high-quality SCs of brain connectivity by using recent advances for reconstructing long-range white matter connections such as local fiber orientation estimation on multi-shell dMRI data with constrained spherical deconvolution, which yields both increased sensitivity to detecting crossing fibers compared with competing methods and the ability to separate signal contributions from different macroscopic tissues, and improvements to streamline tractography such as anatomically constrained tractography and spherical-deconvolution informed filtering of tractograms, which have increased the biological accuracy of SC creation. Here, we provide step-by-step instructions to creating SCs by using these methods. In addition, intermediate steps of our procedure can be adapted for related analyses, including region of interest-based tractography and quantification of local white matter properties. The associated software MRtrix3 implements the relevant tools for easy application of the protocol, with specific processing tasks deferred to components of the FSL software. The protocol is suitable for users with expertise in dMRI and neuroscience and requires between 2 h and 13 h to complete, depending on the available computational system.