The invention discloses a micro-fluidic chip and a manufacturing method thereof, and belongs to the field of biochemical detection. The method comprises the steps that a preset film is formed on a substrate structure; the preset film is machined into a first tooth-shaped structure; a structural layer is arranged on the substrate structure with the first tooth-shaped structure so as to form a micro-fluidic channel. The tooth-shaped structure is formed on the substrate structure, and the micro-fluidic channel is formed by the tooth-shaped structure, the substrate structure and the structural layer, so that the micro-fluidic chip is formed, and a mold of the tooth-shaped structure does not need to be manufactured in advance. The problems that in the prior art, a tooth-shaped mold needs to be manufactured at first, the preset film comprising the tooth-shaped structure is formed through the mold, and then, the preset film is stripped from the mold, so that the manufacturing process of the micro-fluidic chip is complex, and the manufacturing efficiency is low are solved, and the effects that the manufacturing process of the micro-fluidic chip is simplified and the manufacturing efficiency of the micro-fluidic chip is improved are achieved.