Back ground: The feeding practice of new born has implication for immediate and future health and those children exposed for Prelacteal feeding/PLF/ were highly affected by infections like diarrhea, pneumonia and chronic diseases, such as heart disease & type two diabetes mellitus. Objective: The study was aimed to assess the prevalence and associated factors of Prelacteal feeding/PLF/ practice among mothers of children less than one year in Banja woreda, Awi zone, Northwest Ethiopia. Method: A Community based quantitative cross sectional study was conducted from OctoberDecember/2016 in Banja woreda and a total of 481 mothers with under one-year age children were participated in the study. A pre-tested questioner was used to collect the data. Bivariate and multivariate logistic regression model was fitted to identify factors associated with PLF practice. Result: The prevalence of PLF was found to be 12.3%. mothers who gave birth at home were 4.6 times more likely to practice prelacteal feeding as compared to mothers who delivered at health institutions (AOR = 4.6, 95 % CI 2.2, 9.6) and the odds of PLF were 2.8 times (AOR=8.2, 95% CI 5.145, 13.404) more likely among mothers who didn’t have knowledge on the risk of PLF than mothers who know the risk of PLF. Conclusion and recommendation: The prevalence of Prelacteal feeding was relatively low in the study area. Not having knowledge on risk of PLF and home delivery were found to be determinant factors associated with the magnitude of PLF. Thus, Social Behavioral Change Communication/SBCC/ on promotion of institutional delivery and IYCF practice should have to be strengthened at all level.