Nurse preceptors play an important role in supporting newly qualified nurses during transition periods. However, limited attention is given to the needs and experience of nurse preceptors with expected responsibilities. This study aimed to examine the perceived needs of nurse preceptors in three public acute hospitals by using a sequential mixed method approach conducted between March and August 2017. A questionnaire that comprised socio-demographic data, Clinical Teaching Behaviour Inventory (CTBI), and RN Preceptor Learning Needs Assessment, was distributed to all nurse preceptors. Semi-structured qualitative interviews were conducted with a purposive sample of 10 informants to complement the quantitative findings. We received 260 completed questionnaires, giving a response rate of 78.8%. The highest mean CTBI domain score was “Using appropriate teaching strategies” (Mean = 3.65, SD = 0.56), whereas the lowest was “Providing feedback and evaluation” (Mean = 3.51, SD = 0.60). The top five topics identified as the most important in nurse preceptor training were critical thinking, prioritising, teaching techniques, conflict management and teamwork. Qualitative findings revealed that the informants experienced tension with their dual roles and strained relationships with co-workers. The expectations of the informants for support were recognition from management level and highlighting coaching tactics, reciprocal learning and collegiate support.