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[Cu(L)] (1) and [Ni(L)] (2) [L = N-2-hydroxyacetophenon-N′-2-hydroxynaphthaldehyde-1,2 phenylenediimine)], Schiff base complexes have been successfully synthesized. Single crystal X-ray crystallography, Fourier Transform infrared spectroscopy (FT-IR), NMR spectroscopy and elemental analysis (EA) have been used to characterize the complexes. Complex 1 shows unsymmetrical ligand coordination to copper (II) ion in a tetradentate fashion through ONNO donor atoms according to single crystal X-ray patterns. Spectroscopic methods have been used to study the interactions between the synthesized Schiff base complexes and Calf Thymus DNA (CT-DNA) and Bovine Serum Albumin (BSA) under physiological conditions. UV–vis absorption, fluorescence, and circular dichroism (CD) spectroscopy and thermal denaturation measurements were used in the investigation of DNA binding properties of these two …