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This study investigated the effect of the bonding agent application method on the dentin bond durability of a two-step adhesive utilizing a universal-adhesive-derived primer. The bonding durability of a universal adhesive was compared with those of two conventional two-step adhesives by shear bond strength testing after thermal cycling. The primer was applied (with or without phosphoric acid pre-etching), and the specimens were divided into three groups based on the bonding agent application method used as follows: (i) strong air-flow application, (ii) gentle air-flow application, and (iii) gentle air-flow + second bonding application. The shear bond strength was determined after thermal cycling. All the factors (bonding agent application method, adhesive system, and storage condition) significantly influenced the dentin shear bond strength both with and without phosphoric acid pre-etching. The specimens exposed to gentle air-flow showed a higher shear bond strength than did those exposed to strong air-flow and gentle air-flow + second bonding. The bond durability of the tested adhesive systems was influenced by the bonding agent application method, and this trend was material dependent. The G2-Bond Universal exhibited the same or greater dentin bond durability to the other two-step adhesive systems.