Many diffusion tensor tractography (DTT) studies have reported on the topography of transcallosal fibers (TCF). However, little detailed anatomical information on TCF that can be easily applied for clinical purposes is known. Using probabilistic DTT, we attempted to determine the anatomical location of the TCF for motor and sensory function in the human brain. A total of 51 healthy subjects were recruited for this study. Diffusion tensor images (DTIs) were obtained at 1.5 T, and four TCF for the premotor cortex (PMC), the primary motor cortex (M1) for hand and leg, and the primary somatosensory cortex (S1) were obtained using FMRIB software. Locations of the TCF were defined as the highest probabilistic location on the midsagittal slice of the corpus callosum. We measured distances between the most anterior and posterior points of the corpus callosum. The relative mean distances of the highest probabilistic location for the precentral knob PMC (Brodmann area 6 anterior to the precentral knob), hand M1, leg M1, and precentral knob S1 (postcentral gyrus posterior to the precentral knob) TCF were 48.99%, 59.78%, 67.93%, and 73,48% from the most anterior point of the CC, respectively. According to our findings, the precentral knob PMC, hand M1, leg M1, and precentral knob S1 TCF were located at the anterior body, posterior body, posterior body, and isthmus according to Witelson's classification, respectively.