Background: G6PD deficiency is among the leading causes of neonatal hyperbilirubinemia and kernicterus. Following a 2022 New York State Department of Health recommendation to test high-risk neonates, we implemented universal G6PD deficiency screening in the well-baby nursery. Objectives: We aimed to increase the proportion of infants screened from 0% to more than 75% within 6 months and to describe the prevalence of G6PD deficiency and the early outcomes of affected neonates. Methods: This quality improvement (QI) initiative, guided by the Model for Improvement, included a cross-sectional analysis of screening yield and early neonatal outcomes. A statistical process control p-chart tracked monthly screening. Outcomes were compared between screen-positive and screen-negative infants using the Fisher exact test. Results: Screening rose from 0% to a sustained mean of 81.1%, exceeding the 75% aim. Of 580 screened neonates, 52 (9.0%) screened positive for G6PD deficiency. Screen-positive infants were more likely than screen-negative infants to undergo repeat serum bilirubin testing (38.5% vs. 19.3%; p = 0.002) and to reach a peak bilirubin above 10 mg/dL (30.8% vs. 10.2%; p < 0.001). A higher rate of readmission for phototherapy was also observed (5.8% vs. 0.9%; p = 0.028), though based on few events. Phototherapy during the birth hospitalization, IVIG, and exchange transfusion did not differ. Conclusions: Universal G6PD screening was feasibly implemented and sustained in a high-risk well-baby nursery through routine workflow changes, without additional phlebotomy. Nearly 1 in 11 screened neonates tested positive for G6PD deficiency.