ObjectiveThis study aimed to assess mother’s knowledge on neonatal jaundice (NNJ) and its associated factors in northern Ethiopia.DesignFacility-based cross-sectional study.SettingReferral hospitals in Amhara region, Ethiopia.ParticipantsThe study was done among 380 mothers selected using a systematic random sampling technique at referral hospitals in the Amhara region, northern Ethiopia, from 1 March 2019 to 30 July 2019.Primary outcomeMother’s knowledge on NNJ, modelled using binary logistic regression.Secondary outcomeFactors associated with mother’s knowledge about NNJResultsThis study revealed that the overall mother’s knowledge on NNJ was 39.2%. Having favourable attitude (AOR=2.05, 95% CI: 1.25 to 3.34), having a history of NNJ in previous children (AOR=7.51, 95% CI: 3.12 to 18.05), having a history of NNJ in the current child (AOR=1.97, 95% CI: 1.19 to 3.27), antenatal care (ANC) follow-up (AOR=3.85, 95% CI: 1.24 to 14.55) and resided in the urban area (AOR=2.63, 95% CI: 1.32 to 5.25) were factors significantly associated with mother’s knowledge on NNJ.ConclusionsThis study showed that mothers' knowledge on NNJ was low. The attitude of mothers on NNJ, previous and current child history of NNJ, ANC follow-up and residence were variables that had a significant association with mother’s knowledge on NNJ. Enhancing NNJ educational programmes targeting mothers in raising awareness on the prevention of NNJ in the population are the recommended interventions.