Nanosilver, being the most prominent nanoproduct has diverse bio-medical applications and hence the effects associated with their exposure need to be investigated in detail. The interaction of metal nanoparticles with DNA has become a matter of interest, as their effect on structural integrity, synthesis and replication could be explored through it. Present work aims at the facile synthesis and characterization of spherical silver nanoparticles (AgNPs) using Epipremnum aureum leaves extract. Nanoparticles were characterized using UV-Visible spectroscopy, Transmission Electron Microscopy (TEM), High Resolution X-ray Diffraction (HR-XRD) and Dynamic Light Scattering (DLS) studies. The interaction of AgNPs with Calf thymus DNA (CT-DNA) was investigated using different spectroscopic techniques like UV-Visible spectroscopy, UV-thermal melting, Circular Dichroism and fluorescence spectroscopic studies. Fluorescence results suggest van der Waals and H-bonding interactions, which are predominantly responsible for the interaction of AgNPs with CT-DNA. Circular dichroism and thermal melting studies are pointing towards the groove binding of AgNPs to CT-DNA. DNA duplex destabilization was confirmed by the decreased thermal melting temperature of CT-DNA on addition of AgNPs. Present study might open up new vistas for the study of unusual kind of DNA binders, which can destabilize DNA and may further be used for various biomedical applications.