In a world filled with big data, mathematical models, and statistics, the development of strong quantitative skills is becoming increasingly critical for modern biologists. Teachers in this field must understand how students acquire quantitative skills and explore barriers experienced by students when developing these skills. In this study, we examine the interrelationships among gender, grit, and math confidence for student performance on a pre-post quantitative skills assessment and overall performance in an undergraduate biology course. Here, we show that females significantly underperformed relative to males on a quantitative skills assessment at the start of term. However, females showed significantly higher gains over the semester, such that the gender gap in performance was nearly eliminated by the end of the semester. Math confidence plays an important role in the performance on both the pre and post quantitative skills assessments and overall performance in the course. The effect of grit on student performance, however, is mediated by a student's math confidence; as math confidence increases, the positive effect of grit decreases. Consequently, the positive impact of a student's grittiness is observed most strongly for those students with low math confidence. We also found grit to be positively associated with the midterm score and the final grade in the course. Given the relationships established in this study among gender, grit, and math confidence, we provide "instructor actions" from the literature that can be applied in the classroom to promote the development of quantitative skills in light of our findings.