Palatal injections are classified among the most painful dental injections. Therefore, it was necessary to find alternatives to this painful injection to enhance children's cooperation. It was mentioned in the dental literature that EMLA cream was used as a possible alternative to conventional injections, but its anesthetic effect was controversial, so it was useful to study the effect of chemical enhancers to rise the effectiveness of this cream. This trial aimed to evaluate for the first time the anesthetic efficiency and the pain related to the application of the mixture of the chemical permeability enhancer “dimethyl sulfoxide” with EMLA cream in comparison to EMLA cream only and conventional palatal injections in children. This randomized controlled trial included 90 children aged between 7- 11 years with one primary upper molar indicated to extraction. Participants were randomly assigned into three groups to anesthetize the palatal mucosa; G1 (control): conventional palatal anesthesia, G2: EMLA cream only, and G3: EMLA with DMSO. Then, the buccal mucosa was anesthetized with conventional anesthesia, and then the molar was extracted. The pain was assessed by the FLACC scale during palatal probing, and extraction. Kruskal-Wallis test showed significant differences between groups in FLACC scores of the palatal probing stage (P = 0.013). Mann-Whitney U showed a significant difference in pairwise comparisons between G1 and G2 during palatal probing (P = 0.035). The application of EMLA cream for 3 minutes is not sufficient to achieve palatal anesthesia, while the mixture of dimethyl sulfoxide with EMLA cream can be an efficient alternative to conventional palatal injections. Dental injection causes fear and anxiety in children, which leads to a change in their behavior and a loss of cooperation, so it is important to find alternative ways to perform good anesthesia to make dealing with children easier and more acceptable.