© 2022 Scandinavian Division of the International Association for Dental Research. Published by John Wiley & Sons Ltd.This study aimed to evaluate the effect of non-thermal atmospheric pressure plasma on the bond strength of a universal adhesive used in etch-and-rinse mode. Dentin surfaces were etched with phosphoric acid and samples were divided into groups exposed to either dry bonding, plasma-dried bonding, plasma-dried and rewetted bonding, or wet bonding (n = 10). Dentin surfaces of the plasma-dried specimens were treated with a plasma jet before the adhesive procedure. After application, composite blocks were built, and specimens were subjected to micro-tensile bond strength testing after 24 h and after 10,000 thermal cycles. The hybrid layer formation was evaluated by micro-Raman spectral analysis; the resin–dentin interface was analyzed by scanning electron microscopy. One-way ANOVA and Tukey's post hoc multiple comparison tests were used to statistically analyze the data. The bond strength values of the plasma-dried bonding groups were statistically higher than the non-plasma-treated groups both before and after aging. After the thermal cycles, bond strength values decreased significantly only in the wet bonding group. Micro-Raman spectral analysis revealed that plasma-drying increased adhesive penetration, especially hydrophobic monomer infiltration. This may increase the mechanical properties and durability of the resin–dentin interface, provide long-term stability, and improve the polymerization rate of the adhesive layer.