Disclosed are a manufacturing method of an anti glare layer and a display panel. The method comprises: preparing a spray solution comprising sprayed particles, each having a shell structure; spraying the spray solution uniformly onto the glass substrate by a spraying device to form an arrangement structure on the glass substrate; spraying an acidic liquid on the arrangement structure to remove the shell structures of the sprayed particles each having the shell structure to form gaps among the spray particles; implementing an annealing treatment to the sprayed particles to form the anti glare layer. Thus, the light diffraction effect can be reduce or even eliminated to improve the picture quality of the display for promoting the comfort of the user viewing.