Background The establishment of human identity has always been a concern after mass disasters, and the role of odontology in forensics has greatly evolved. Nowadays, palatal rugoscopy is a widely used method in the recognition of human identity due to its uniqueness in the course, direction, length, form, position, and enduring nature against disintegration. Its easy applicability, cost-effectiveness, and prompt results can be applied to festering, scorched bodies and in the absence of missing upper limbs and fingers. This study was undertaken to evaluate palatal rugoscopy as a tool to recognize human identity and gender between two different ethnic populations. Methodology A cross-sectional comparative study was conducted in the Department of Oral Maxillofacial Surgery and Diagnostic Sciences, Riyadh Elm University, Riyadh, Saudi Arabia, to recognize ethnicity and gender among the Saudi and Kuwaiti populations. A total of 364 participants were selected from the outpatient department between September 2022 and December 2022. All 364 participants were distributed into two groups after the application of inclusion and exclusion criteria. After obtaining informed consent from all study participants, study models were prepared for final interpretation. The outlining of rugae was done with the help of a sharp graphite pencil, and the assessment of various parameters, including total number, length, direction, unification, and shape of rugae, was done. A comparison was made between the two populations. SPSS version 26.0 (IBM Corp., Armonk, NY, USA) was employed to assess variations in the mean values of both the total number and the distribution of rugae on the right and left sides across different ethnic groups and genders. Results In this study, a total of 364 participants were included. Of the total participants, 184 were Saudis and 180 were Kuwaitis, with 188 males and 176 females. A comparative evaluation of rugae among ethnic groups showed that Saudi participants had a mean number of 8.92 ± 0.660 palatal rugae, whereas in Kuwaiti participants it was 8.68 ± 0.649 (p = 0.001). When rugae length was assessed between genders among Saudi participants, it was found that the majority of participants had primary rugae with a length of more than 5 mm, and the difference was statistically significant (p = 0.002). The majority of Saudi males had forwardly positioned rugae, while the majority of Saudi females had more backwardly placed rugae. When rugae length was assessed in males and females among Kuwaiti participants, it was found that males had more primary rugae than females. This study found that the majority of Kuwaiti males had a wavy shape of rugae, while the majority of Kuwaiti females had more straight rugae. Conclusions This study concluded that among the two ethnic groups, the total number, length, direction, unification, and shape of rugae were different between genders, with significant differences in some parameters. Therefore, palatal rugoscopy might be useful as a tool to recognize gender and ethnicity and may provide better results when used as an additional tool along with other dependable forensic tools., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2024, Abdul et al.)