The hydroxyapatite (HA) formation on the DNA molecules in SBF was examined. After immersion for four weeks in SBF at 36.5 °C, the HA crystallites of ~1-14 μm in diameter grew on the surface of DNA molecules. Various morphologies were successfully observed through scanning electron microscopy analysis. The Ca/P mol ratio (1.1-1.5) in HA was estimated by energy dispersive X-ray analysis. Original peaks of both of DNA and HA were characterized by Fourier transform infrared spectroscopy. The molecular orbital computer simulation has been used to probe the interaction of DNA with two charge-balancing ions, i.e., CaOH(+) and CaH2PO4(+). The adsorption enthalpy of the two ions on ds-DNA and/or ss-DNA having large negative value (~ -60 kcal/mol per charge-balancing ion) was the evidence for the interface in mineralization of HA in SBF.