The aim of this study was to evaluate the microtensile bond strength (μTBS) of one layer or two layer applications of single bond (SB) and prime & bond (PB) which prepared with erbium: Yttrium aluminum-garnet (Er:YAG) laser, potassium titanyl phosphate (KTP) laser, and acid-etched dentin. Various surface pretreatments have been developed to have better adhesion to dentine. However, the effects of laser-etch and acid-etch on adhesion with various applications of bonding agents have not yet been achieved. Twenty-four human third molar teeth were used in this study. One third of the teeth were removed; then the teeth divided into three groups and two subgroups randomly. After Er:YAG laser, KTP laser, and acid-etch applied to dentine, SB and PB applied one or two layer for the subgroups. Waiting in distilled water for 24 h, the teeth were sectioned to sticks and the sticks were stressed in tension until failure using a microtensile testing machine. The data were collected in SPSS for statically analysis with three-way ANOVA and independent sample t-tests at the confi dence interval of 95%. The highest μTBS values were in Er:YAG laser and PB group in one-layer application. The Er:YAG laser can be useful to obtain high μTBS values. Applying a second consecutive coat of adhesive showed decreased μTBS values and KTP laser was not effective to obtain an increased μTBS values.