Aims: The demand for blood and blood products in sub-Saharan Africa is occasioned by tropical infectious diseases and obstetric complications that cause anemia. We therefore need a well-organized blood transfusion service with sufficient resources to process blood and blood products, especially in the Western Kenya, where co-infections of tropical diseases are rampant. This study was aimed at determining the blood deficiency and challenges experienced by Eldoret Regional Blood Transfusion Center (ERBTC). Methods: A cross-sectional descriptive design was used to conduct the study. All the ERBTC staffs, donor records, and blood requisition and dispatch registers were studied. Self-administered questionnaires and data collection abstraction forms were used to collect data. The extracted data from ERBTC registers included those on demand and supply mismatches and blood discards. The obtained data were entered into Microsoft Excel and analyzed using descriptive statistics. Results: A total of 16 staffs, 230 donor records, and 9612 units of requested blood were reviewed. The ERBTC reported many challenges ranging from understaffing, limited funding, insufficient equipment, and irregular reagents and other laboratory supplies. During the study period, the blood bank only managed to supply 4740 units of blood against a demand of 9612 units, thus occasioning a 50.7% deficit. A discard rate of 7.8% after the screening was also reported due to insufficient volumes after collection and transfusion transmissible infections. However, there were no wastages during the study period. Conclusion: The main challenges experienced by ERBTC were understaffing, insufficient funding, limited equipment, frequent reagent outages, and discards of unsuitable blood, resulting in very high deficits. We recommend hiring staff, increased funding, acquiring modern equipment and reagents, and recruiting known regular blood donors to alleviate frequent shortage.