The present application provides a flexible substrate and a manufacturing method of the flexible substrate. The flexible substrate includes a first flexible layer, a barrier layer, a second flexible layer, and a buffer layer. The barrier layer is disposed on a surface of the first flexible layer, the barrier layer includes a first bending resistant structure disposed therein, the second flexible layer is disposed on a surface of the barrier layer, the buffer layer is disposed on a surface of the second flexible layer, and the buffer layer includes a second bending resistant structure disposed therein.