OBJECTIVE: To compare two different doses of lidocaine used for periprostatic nerve block on pain perception during transrectal ultrasound (TRUS) guided prostate biopsy. MATERIAL AND METHODS: A total of 288 patients with elevated prostate specific antigen (PSA) levels and/or abnormal digital rectal examination who underwent TRUS-guided prostate biopsy were included in the study. The patients were divided into 3 groups: Group 1 (n=103) prostate biopsy were performed after administering perianal intrarectal application of 10 mL 2% lidocaine gel, Group 2 (n=98) 2 mL of 2% lidocaine injection on each side following rectal installation of lidocaine gel and Group 3 (n=87) 4 mL of 2% lidocaine injection on each side after rectal instillation of lidocaine gel. Patients' pain scores during biopsy procedure were reported using visual analogue score (VAS). Independent sample t test, ANOVA test and Tukey test were used for statistical evaluation. RESULTS: The mean age, prostate volume and PSA level were 65.6±8.4 years, 58.2±34.8 mL, and 11.8±3.4 ng/mL respectively. There were no statistically significant differences in baseline characteristics between the groups. The mean VAS scores were 2.4±1.8 in Group 1, 2.5±1.9 in Group 2 and 1.6±1.6 in Group 3. Patients in Group 3, reported significant pain reduction compared with patients in Groups 1 and 2 (p=0.002, and 0.001, respectively). However, there was no statistically significant difference in VAS scores between Groups 1 and 2 (p=0.815). CONCLUSION: According to our results we recommend the use of perianal intrarectal lidocain gel application, and periprostatic nerve block with injection of 4 ml 2% lidocaine per side combination in TRUS-guided prostate biopsies. Further large-scale randomized control studies are needed to validate these finding.