There is provided a method of forming a nitride semiconductor layer, including the steps of firstly providing a substrate on which a patterned epitaxy layer with a pier structure is formed. A protective layer is then formed on the patterned epitaxy layer, exposing a top surface of the pier structure. Next, a nitride semiconductor layer is formed over the patterned epitaxy layer connected to the nitride semiconductor layer through the pier structure, wherein the nitride semiconductor layer, the pier structure, and the patterned epitaxy layer together form a space exposing a bottom surface of the nitride semiconductor layer. Thereafter, a weakening process is performed to remove a portion of the bottom surface of the nitride semiconductor layer and to weaken a connection point between the top surface of the pier structure and the nitride semiconductor layer. Finally, the substrate is separated from the nitride semiconductor layer through the connection point.