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Background: The diagnostic efficacy regarding prostate cancer (PC) detection by manually operated in-bore magnetic resonance imaging (MRI) targeted prostate biopsy (MO-MRGB) versus robot-assisted in-bore MRI targeted prostate biopsy (RA-MRGB) is lacking evidence., Objective: We hypothesized that the detection rates (DRs) for PC of MO-MRGB and RA-MRGB were similar and aimed to compare these., Design Setting and Participants: We prospectively included all patients who received in-bore MRI targeted prostate biopsy (MRGB) of the prostate in the Central Denmark Region from August 2014 to February 2020. From August 2014, MO-MRGB was used, and from March 2018, RA-MRGB was preferred. Referral to in-bore MRGB was based on multiparametric MRI (mpMRI)., Outcome Measurements and Statistical Analysis: We compared PC DRs of MO-MRGB and RA-MRGB with Pearson's chi-square test. We made three binary regression models and calculated the risk difference (RD) of PC between the in-bore MRGB systems., Results and Limitations: A total of 3107 patients were referred to mpMRI, and 884 (28%) patients went on to receive in-bore MRGB. The MO-MRGB and RA-MRGB systems were used in 505 (57%) and 379 (43%) patients, respectively. Taking clinically relevant covariates into account, we found no statistically significant difference in PC DRs between MO-MRGB and RA-MRGB (72% vs 73%, RD 1%, 95% confidence interval -4% to 7%, p  = 0.6). The main limitation was a shift in population characteristics., Conclusions: We did not see evidence of an effect on the DR or the RD for PC when we compared MO-MRGB with RA-MRGB. Cost effectiveness should be considered carefully when choosing the MRGB system., Patient Summary: We compared two magnetic resonance imaging guided prostate tissue sampling systems regarding prostate cancer (PC) detection. One system was manually operated, and the other system was robot assisted. Comparing the systems, we found no evidence of a difference in their ability to detect PC., (© 2022 The Author(s).)