A method of manufacturing a display substrate includes forming a first metallic pattern including gate and storage conductors and a gate electrode of a switching device on a base substrate, forming a gate insulation layer, forming a second metallic pattern and a channel portion including a source line, source and drain electrodes of the switching device, forming a passivation layer and a photoresist film on the second metallic pattern, patterning the photoresist film to form a first pattern portion corresponding to the gate and source conductors and the switching device, and a second pattern portion formed on the storage line, etching the passivation layer and the gate insulation layer, and forming a pixel electrode using the first pattern portion. Therefore, excessive etching of the stepped portion may be prevented, so that a short-circuit defect between a metallic pattern and a pixel electrode may be prevented.