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Abstract Objective The aim of this study was to assess nurses’ Knowledge, Attitude and Associated Factors towards end of life care in Amhara Referral Hospitals, Northwest Ethiopia, 2017. Results A total of 331 participants were included with a response rate of 93.2%. From these, 129 (39.0%) of them had good knowledge and 234 (70.7%) had favorable attitude towards end of life care. Being Bachelor of Science holder and above in nursing (AOR = 4.261, 95% CI 1.524–11.912), working in Emergency department (AOR = 4.911, 95% CI 1.796–13.426), having daily experience of caring for chronically ill patients (AOR = 2.764, 95% CI 1.366–5.591) and taking training on end of life care (AOR = 10.269, 95% CI 4.730–22.296) were significantly associated with good knowledge of nurses towards end of life care. On the other hand, having 6–10 years of working experience in nursing (AOR = 2.199, 95% CI 1.147–4.215), being trained in end of life care (AOR = 3.027, 95% CI 1.285–7.13), Bachelor of Science holder and above in nursing (AOR = 4.414, 95% CI 2.230–8.738) were found to be statistically significant with more positive attitude of nurses towards end of life care.