The goal of our study was to evaluate the impact of the interval between prostate biopsy and magnetic resonance imaging (MRI) on the accuracy of simple tumor localization, which is essential information that enables nerve-sparing surgery. We also sought to determine the optimal timing of a post-biopsy MRI. A total of 184 patients who had undergone MRI before radical prostatectomy at an institution without a predetermined schedule for MRI after a prostate biopsy were enrolled. The mean interval from the biopsy to the MRI was 30.8 ± 18.6 days. The accuracy of the MRI for simplified tumor location (right, left, bilateral and none) was 44.6%. In the group with discordant pathologic and MRI findings, the most common reason recorded was ‘MRI predicted a unilateral lesion, but pathology revealed bilateral lesions’ (58.3%), followed by ‘MRI predicted no lesion, but pathology revealed the presence of a lesion’ (32.0%). Multivariable analysis showed that the discordant group had a shorter interval (25.0 ± 14.3 vs 38.1 ± 20.6 days, P < 0.01) preceding the MRI and a higher rate of hemorrhage as observed by MRI (80.4% vs 54.8%, P < 0.01) in comparison with the accordant group. In receiver operating characteristics analysis, the area under the curve of the MRI interval in accurate prediction of the tumor location was 0.707 (P < 0.001). At the MRI interval's cutoff of 28.5 days, the sensitivity was 73.2% and the specificity was 63.7%. When the MRI was performed within 28 days, the accumulated accuracy was only 26.1% (23/88); however, when it was performed after 28 days, the reversely accumulated accuracy was 61.5% (59/96). These data support a waiting period of at least 4 weeks after a biopsy before performing an MRI for the purposes of surgical refinement.