Most of deep geological engineered structures, such as rock caverns, nuclear waste disposal repositories, metro rail tunnels, multi-layer underground parking, are constructed within hard crystalline rocks because of their high quality and low matrix permeability. In such rocks, fluid flows mainly through fractures. Quantification of fractures along with the behavior of the fluid flow through them, at different scales, becomes quite important. Earlier studies have revealed the influence of sample size on the confining stress–permeability relationship and it has been demonstrated that permeability of the fractured rock mass decreases with an increase in sample size. However, most of the researchers have employed numerical simulations to model fluid flow through the fracture/fracture network, or laboratory investigations on intact rock samples with diameter ranging between 38 mm and 45 cm and the diameter-to-length ratio of 1:2 using different experimental methods. Also, the confining stress, σ3, has been considered to be less than 30 MPa and the effect of fracture roughness has been ignored. In the present study, an extension of the previous studies on “laboratory simulation of flow through single fractured granite” was conducted, in which consistent fluid flow experiments were performed on cylindrical samples of granitoids of two different sizes (38 mm and 54 mm in diameters), containing a “rough walled single fracture”. These experiments were performed under varied confining pressure (σ3 = 5–40 MPa), fluid pressure (fp ≤ 25 MPa), and fracture roughness. The results indicate that a nonlinear relationship exists between the discharge, Q, and the effective confining pressure, σeff., and Q decreases with an increase in σeff.. Also, the effects of sample size and fracture roughness do not persist when σeff. ≥ 20 MPa. It is expected that such a study will be quite useful in correlating and extrapolating the laboratory scale investigations to in-situ scale and further improving theoretical/numerical models associated with fluid flow through rock masses.