There are provided a substrate with a reflective layer and an EUV mask blank, which can prevent particles from adhering to a surface of the reflective layer or an absorbing layer, or into a reflective layer or an absorbing layer during formation thereof by eliminating electrical connection between a film formed on a front surface of the substrate and a film formed on a rear surface of the substrate.A substrate with a reflective layer, which is usable to fabricate a reflective mask blank for EUV lithography, comprising a chucking layer formed on a rear surface opposite a surface with the reflective layer formed thereon, the chucking layer serving to chuck and support the substrate by an electrostatic chuck, wherein the reflective layer has no electrical connection to the chucking layer.