Aim: The aim of this research was to identify the factors that Registered Nurses (RNs) in Australia perceived as impacting on their clinical practice in the ART industry. Method: A Grounded Theory methodology guided the conduct of this study. After gaining ethical clearance, fifteen in-depth individual interviews were undertaken to identify and explore ART Nurses’ perceptions in relation to their clinical practice. A constant comparative approach was used throughout data analysis. Results: The participants identified context-specific elements and a series of intrinsic and extrinsic factors that they believed influenced their clinical practice. RN attributes (both personal and professional); knowledge; skills; and patient care activities were shown to be the key context-specific elements. The intrinsic and extrinsic factors (positive and negative) that were shown to impact on ART nursing practice included: communication; support; teamwork; patient issues; and time. Conclusion: This translational research identified the central role of the ART nurses as they ensure that a ‘healthy start’ is established and maintained throughout the patient journey. Intrinsic and extrinsic factors that impacted on practice were highlighted and will inform practice and advice on educational requirements. Findings from this in-depth qualitative study are relevant to nurses and health service providers in the ART community.