A display substrate and a manufacturing method thereof, and a display device are provided. The display substrate includes a base substrate, a pixel drive layer, a light-emitting device, an encapsulation layer, a first insulation layer, and a covering layer. The base substrate includes a display region and a peripheral region, the peripheral region includes a first peripheral region and a second peripheral region. The first insulation layer is in the second peripheral region, and includes a first notch, a side edge of the first notch away from the display region overlaps with the edge of the base substrate. The covering layer is at least partially filled in the first notch, and an orthographic projection of the covering layer on the base substrate at least partially overlaps with an orthographic projection of the first notch on the base substrate.