Purpose: We aimed to evaluate the radiation exposure to patients undergoing positron emission tomography/computed tomography (PET/CT)-guided biopsies. Materials and Methods: Patients undergoing PET/CT-guided biopsy were recruited prospectively from October 2019 to April 2020. PET/CT-guided biopsy from a tracer avid site was done using an automated-robotic-arm 1 h after intravenous injection of F-18-fluorodeoxyglucose (FDG) (2-5 mCi) or Ga-68-PSMA (1–4 mCi). Regional CT-images were acquired for biopsy planning and confirmation of needle placement. The internal radiation exposure due to the PET component was estimated using the value of activity injected and dose-coefficient for FDG and PSMA. The external radiation exposure due to the CT component was estimated using the value of dose length product and organ coefficients conversion factor. The total effective dose during the procedure was calculated by adding exposure due to both CT and PET components. Percentage contribution from CT and PET component to total effective dose was compared using a paired t-test. Results: A total of 101 patients (76 males) were recruited for PET/CT-guided biopsy using FDG (n = 79) and PSMA (n = 22). The mean effective-dose due to PET and CT components and total effective-dose was 2.49 ± 1.02 mSv, 2.35 ± 1.03 mSv and 4.83 ± 1.90 mSv, respectively, for FDG-guided procedures and 1.60 ± 0.57 mSv, 3.06 ± 1.36 mSv, and 4.66 ± 1.37 mSv for Ga-68-PSMA-guided procedures. The percentage contribution of PET and CT in total effective-dose was comparable in F-18-FDG and Ga-68-PSMA PET/CT-guided biopsy procedures; however, for Ga-68-PSMA PET/CT-biopsies, CT contributed a higher radiation dose than PET component. Conclusion: PET/CT-guided biopsy is a safe interventional procedure, and radiation exposure to the patients was less than routine whole-body PET/CT-imaging.