A manufacturing method of a glass substrate is provided, in which a mask including a light-blocking area, a transparent area and a partial-transparent area is adopted. The partial-transparent area protrudes from edges of the light-blocking area to admit some of the UV rays to pass through. In addition, a glass substrate manufactured with the method is also disclosed. By arranging the partial-transparent area on the edges of the light-blocking area, the mask is formed with a slope having a small angle after a lithography process. As such, in an etching process, an edge of a thin film is formed with a slope having a small angle, which contributes to the formation of a second thin film. The thin films are prevented from being fragmented around the slope and the ITO layer is also prevented from fragmented around the periphery of the through hole.