The embodiments of the present disclosure provide a display substrate, a display panel and a manufacturing method thereof. The display substrate includes a body portion and a to-be-cut portion arranged on at least one side of the body portion, and the body portion and the to-be-cut portion are in a same plane and connected to have a one-piece structure; the display substrate further includes a plurality of test electrodes, some test electrodes are arranged on the body portion, and the other test electrodes are arranged on the to-be-cut portion. The display panel includes the body portion in the display substrate.