Ionic Liquids having great green properties such as low vapour pressure, tunable viscosity, tunable electrochemical window and wide liquid range which attract the attention for their study. In this course of project the introduction of Ionic Liquids with DNA-Ligand complex was applied, and the effect of ionic liquids on DNA-Ligands interaction were studied by fluorescence spectrophotometry titration. For this purpose Calf-thymus (Ct-DNA) and EtBr was taken under study, a UV-Vis Abs spectroscopic titration at different salt condition were studied and then two different ILs [bmim]Cl and [bpip]NtF2 were introduced with EtBr to study their effect on Ct-DNA-EtBr interaction. It was observed that there was a gradual decrease in binding affinity of EtBr with Ct-DNA as concentration of ILs was being increased. This result coincides with the result as observed with the event when binding affinity was measured at different salt concentration of phosphate Buffer in traditional methods.