Background and Objectives: Uttarakhand, a state in northern India, is home to various local communities, including the Garhwali, Kumaoni, and Jaunsari. In addition, individuals from neighboring countries such as Nepal and Tibet reside in Uttarakhand for many years, resulting in a diverse population. However, currently, there is no regional registry for rare blood donors in this state. The aim of this study was to develop a rare donor registry in Uttarakhand which could in future become a part of the national rare blood donor registry. Methods: During the routine donation process, samples were taken from randomly selected O-typed blood donors who are residents of Uttarakhand. Seventeen monoclonal antisera included Rh (anti-D, anti-C, anti-c, anti-E, and anti-e), Duffy (anti-Fya and anti-Fyb), Kell (anti-K and anti-k), Kidd (anti-Jka and anti-Jkb), Lewis (anti-Lea and anti-Leb), M, N, S, and s (Ortho Diagnostics Pvt Ltd, Mumbai, India). Furthermore, four polyclonal antisera from our serum repository, Diego (anti-Dia), Indian (anti-Ina), Rh (anti-Cw), and anti-Gp-Mur were used for phenotyping. The study was financially aided by the Uttarakhand State Council of Science and Technology (UCOST), Dehradun, Uttarakhand, Government of India. Results: Over a period of 16 months (November 2021–February 2023), out of a total of 17,004 blood donations, O blood type donors accounted for 29.9% (5085/17,004) of the donors screened. Among these, 7.3% (374/5085) of the donors were from the interior and plain areas of Uttarakhand. Our study revealed a high prevalence of k and e antigens, with 96.7% and 92.5%, respectively. The least prevalent antigen was Rh (Cw), observed in only 0.5% of cases. We also identified rare blood antigens such as Diego, Indian, and Gp-Mur and compared our findings with other studies. Furthermore, we identified a donor with an H-deficient phenotype, specifically the Bombay blood group (Oho). Conclusion: We have successfully established a regional registry for rare blood donors in Uttarakhand, which will be valuable for managing complex transfusion scenarios in the future. Further, given Uttarakhand's proximity to China and Nepal, the presence of individuals from these countries aptly connects the fields of anthropology and transfusion medicine.