Background: Hand hygiene is crucial for safe healthcare. Although the use of alcohol hand rubs is encouraged in clinics, there are few studies that focus on the proper use of alcohol hand rubs among nursing students. Objectives: The aim of this study is to evaluate the effectiveness of alcohol-based hand rub application and proper hand washing techniques which influence nursing students' hand hygiene compliance to make recommendations for future practice of hand hygiene training. Design: This cross-sectional study was carried out from May 3/June 3 2016 with the participation of 257 nursing students. Their hand hygiene techniques were analyzed using a UV lamp and an alcohol-based mix marked with fluorescence. Results: Of the participants, 77.0% were first-year students and 55.3% were males. The percentage of skin surface covered by alcohol-based hand rub was 82.0% on both hands. The lowest percentages of skin area covered by fluorescent-labelled hand rub were identified in the metacarpal area near the wrist and thumbs. While there was a difference between points for using proper hand-rub technique on the dorsal and palmar surfaces of the hands (p < 0.01), no difference was determined between the right and left hands (p > 0.05). It was found that the largest fluorescent remains were on fingers after hand-washing, and there was a difference in terms of hand-washing points between right-left hands and dorsal-palmar surfaces (p < 0.01). Conclusions: There is a need to improve nursing students' compliance with hand hygiene. It is suggested that the use of hand-rub among students should be popularized, and new techniques that prevent the frequently omitted areas of the hands should be integrated into the curriculum.