Purpose: The aim of this study was to evaluate the effect of gender and knowledge of color in dentistry on the visual shade-matching ability of dental students with no experience in shade matching and without identification skills. Materials and Methods: A total of 32 color-normal participants, 16 female (F) and 16 male (M), completed all phases of the experiment. The control group did not listen to a 60-minute lecture (FNL = females that did not listen; MNL = males that did not listen); the other group listened to a lecture about color in dentistry (FL = females that listened; ML= males that listened). The Toothguide Training Box (TTB) (VITA Zahnfabrik) was used. The final exam consisted of a total of 15 lightness-chroma-hue tasks. The correct selection of lightness (L*), chroma (C*), and hue (h*) was observed, as was the computed shade-matching score, ΣΔE*ab, for each participant. Mann-Whitney U test was used for statistical analysis of the data (α = .05) (SPSS 22.0 for Windows [IBM]). Results: Gender was found to play an important role in shade matching. The FL group selected L* better (L* = 12.11) in comparison with the MNL group (L* = 11.00), which is not significantly different (P = .19). The FL group selected L* better in comparison with the ML group (L* = 10.57), which is not significantly different (P = .10). The FNL group selected C* statistically significantly better (C* = 9.86) than did the ML group (C* = 8.57) (P = .016). The shade-matching score, ΣΔE*ab, for group FL (ΣΔE*ab = 22.50) and group ML (ΣΔE*ab = 31.79) was marginally statistically significant (P = .06). Conclusion: A 60-minute lecture from the field of color in dentistry has a minimal impact on tooth-shade matching, whereas gender plays an important role.