Aim: Root caries is a common, debilitating condition particularly in the elderly population, which can lead to tooth loss. Evidence shows that green tea has cariostatic effects. Considering the gap of information on green tea varnish, this study aimed to assess the efficacy of green tea varnish in the prevention of root caries. Materials And Methods: This in vitro experimental study was performed on 42 sound premolars. Two layers of acid-resistant nail varnish were applied on root surfaces except for a window of 1 × 4 mm. The teeth were randomly divided into three groups (n = 14). Group I (control) received no intervention. Group II received green tea varnish applied on the roots every 48 hours for 21 days. Group III received green tea varnish every 24 hours for 21 days. Sections of 40 μ thickness were prepared from the center of the window, and the depth of carious lesion was measured in three points with 500 μ distance from each other using polarized light microscope. Data were analyzed using Statistical Package for the Social Sciences (SPSS) version 16 and non-parametric Kruskal-Wallis and Mann-Whitney U tests (α = 0.05).Results: The mean (and standard deviation) depth of carious lesion was 54.30 ± 28.64 (μm), 0, and 0 in groups I, II, and III respectively. Control group showed the highest depth of caries. Groups II and III were not significantly different in this respect, but significant differences were noted in depth of caries between groups I and II and also groups I and III (p < 0.001).Conclusion: Based on the results, the green tea varnish may possess anticariogenic effects on root caries when applied every 24 or 48 hours over a 21-day period. Application of green tea varnish every 24 or 48 hours for 21 days may prevent root caries.Clinical Significance: Application of green tea varnish can prevent root caries.