This protocol provides steps for seating and flowing liquid through a 50 μm, 13mm round membrane containing a hexagonal arrangement of 25nm pores with lattice constant 65nm using a peristaltic pump. Porous, alumina membranes are fragile and pressure is required to pass buffer through the 25nm pores.A peristaltic pump at a low flow rate reproducibly hydrates membranes compared to syringe-based assemblies. Steps in this protocol minimize membrane breaks and damage during membrane placement in the filter adapter assembly, liquid flow through the membrane and membrane retrieval.