This study evaluated the marginal microleakage of five single step adhesives. Class V cavity preparations with occlusal margins in enamel and gingival margins in dentin were prepared on both buccal and lingual surfaces of extracted human molar teeth. Prepared teeth were randomly divided into five groups and restored using one of the single step adhesives and composite resins：Prompt L-Pop/Filtek Z-250 (Group 1), AQ Bond/Metafil CX (Group 2), One-Up Bond F/Palfique Toughwell (Group 3). Futurabond/Admira (Group 4), Xeno III/Spectrum TPH (Group 5). The restored teeth were thermocycled. Microleakage was assessed by dye penetration using 2% methylene blue dye solution. The teeth were bisected buccolingually and evaluated for microleakage under steromicroscope. The data were statistically analysed by Kruskal-Wallis test and Mann-Whitney tests. The results of this study were as follows：1. Microleakage of enamel margins in group 3 was statistically higher than that in groups 1, 2, 4, 5 (p < 0.05). 2. Microleakage of dentin margins in group 1 was statistically higher than that in groups 2, 5, and that in group 3 was statistically higher than that in groups 2, 4, 5 (p < 0.05). 3. Dentin marginal microleakage was higher than enamel marginal microleakage in all experimental groups. In conclusion, Prompt L-Pop showed the least leakage at enamel margin, and AQ Bond showed at dentin margin in this study. Marginal miroleakage in dentin was higher than that in enamel.