The mode of binding of 2-(4-(dimethylamino)styryl)-1-methylpyridinium iodide (DASPMI) with calf thymus DNA as revealed from different steady state and time-resolved emission spectroscopic measurements has been reported in this paper. Fluorescence enhancement of DASPMI and its quenching by potassium iodide (KI) points to groove binding of dye with ct-DNA, rather than intercalation in the ct-DNA helix. An increase in steady state anisotropy and fluorescence lifetime hints at binding with ct-DNA. The value of binding constant from emission and association constant from circular dichroic spectrum also indicates weak binding. The strong dependence on ionic strength or salt in controlling the binding of DASPMI with ct-DNA by electrostatic interaction confirms groove binding. The high semicone angle of DASPMI in ct-DNA certainly rules out the possibility of intercalated bonding. A theoretical modeling shows that the probe is bound to ct-DNA as a crescent with a curvature of 11.35 angstrom, which is the previously known curvature of probe in the minor groove.