95 patients undergoing foot and ankle surgery were randomized to receive either 4% chlorhexidine gluconate followed by 70% isopropyl alcohol while preparing the surgical site, or the same two solutions applied in a reverse order. The purpose of this study was to determine whether the order of application of these two solutions has an effect on bacterial cultures, measured at four separate time points in the perioperative period. Results indicated that when 70% isopropyl alcohol was followed by 4% chlorhexidine gluconate, there were significantly less positive bacterial cultures after draping compared to when the reverse order was used. Apart from this finding, there were no significant differences between groups with respect to time to complete the procedure, the number of positive bacterial cultures before and after skin closure, or the incidence of skin or wound infections after 6 months.