To provide a manufacturing method of a semiconductor device in which, even when the semiconductor device is formed over an SOI substrate which uses a glass substrate, an insulating film and a semiconductor film over the glass substrate are not peeled by stress applied by a conductive film in formation of the conductive film for forming a gate electrode. A semiconductor device is manufactured by the steps of forming a first insulating film over a bond substrate, forming an embrittlement layer by adding ions from a surface of the bond substrate, bonding the bond substrate to a glass substrate with the first insulating film interposed therebetween, separating the bond substrate along the embrittlement layer to form a semiconductor film over the glass substrate with the first insulating film interposed therebetween, removing a peripheral region of the first insulating film and the semiconductor film to expose part of the glass substrate, forming a gate insulating film over and in contact with the semiconductor film and the glass substrate, and forming a stacked conductive film over and in contact with the gate insulating film, in which the stacked conductive film includes a conductive film having a tensile stress and a conductive film having a compressive stress.