This study evaluated the in vitro microleakage of six dentin adhesive systems. Triangle-shaped Class V cavities with coronal margin in enamel and gingival margin in cementum or root dentin were cut in the buccal surfaces of 90 non-carious single-root human teeth. These teeth were randomly assigned into six groups (n = 15) for the evaluation of six different dentin adhesive systems: One Step, Prime & Bond 2.0, Syntac Single, Single Bond, Optibond Solo and Syntac Sprint. The preparations were restored with Degufill Ultra composite and polished using the Enhance system. Each group was randomly divided into three subgroups (n = 5): samples of the first subgroup were immersed in 2% methylene blue solution for seven days; those of the second subgroup remained in a similar solution for 31 days; those of the third subgroup were thermocycled 500x at 5-55 degrees C and immersed in 2% methylene blue for seven days. All 90 teeth were then embedded in methacrylate and bucco-lingually sectioned; the dye penetration was evaluated using an 0-4 ordinal scale. All of the dentin adhesive groups showed minimal leakage at the enamel margins with increased leakage at the gingival margins. Optibond Solo showed the best outcomes among the dentin adhesives tested.