OBJECTIVE: To determine whether the method of specimen preparation after transrectal ultrasound-guided biopsy of the prostate affected core fragmentation, and secondly to determine whether such fragmentation was influenced by the presence of malignancy in the biopsy specimen. METHODS: A prospective randomized study comparing 2 different mechanisms of specimen preparation, wash vs swipe of biopsy cores, with blinded assessment by a specific uropathologist was undertaken. RESULTS: For those cores in which prostate cancer was present, the wash method of specimen preparation resulted in fewer and longer core fragments (P = .005). Those patients in whom cancer was present on biopsy also had a greater degree of biopsy core fragmentation in those cores in which only benign tissue was identified (P = .017). Gleason score did not appear to influence the extent of fragmentation. CONCLUSION: The method of specimen preparation at prostate biopsy influences the degree of fragmentation seen at histologic evaluation. This has the potential to affect the interpretation of biopsy results, including suitability of patients for inclusion in active surveillance protocols. Core fragmentation might be minimized by modification of technique.