In adult mammals, the subventricular zone (SVZ) of the lateral ventricles and the subgranular zone of the dentate gyrus (DG) show ongoing neurogenesis, and multipotent neural stem or progenitor cells (NSCs) in these two regions exhibit different intrinsic properties. However, investigation of the mechanisms underlying such differences has been limited by a lack of efficient methods for isolating NSCs, particularly from the adult DG. Here we describe a protocol that enables us to isolate self-renewing and multipotent NSCs from the SVZ and the DG of the same adult mouse. The protocol involves the microdissection of the SVZ and DG from one adult mouse brain, isolation of NSCs from specific regions and cultivation of NSCs in vitro. The entire procedure takes 2-3 h. As only one mouse is needed for each cell isolation procedure, this protocol will be particularly useful for studies with limited availability of mice, such as mice that contain multiple genetic modifications.