The thermal properties and temperature-responsive nanoparticle formation of poly(N-isopropylacrylamide) grafted with single-stranded DNA (PNIPAAm-g-DNA) were investigated. Copolymerization between nonamer single-stranded DNA with a vinyl group at its 5' terminus (DNA macromonomer) and NIPAAm was carried out so that the DNA macromonomer unit content should be less than 1 mol %. The turbidimetry and differential scanning calorimetry of the copolymer showed that the transition temperature increased and the enthalpy change of the phase transition decreased with increasing DNA macromonomer content in the copolymers, indicating that the DNA macromonomer behaves as a hydrophilic part in the copolymer and that the hydrophilicity is greater than that of sodium styrenesulfonate. Above the phase transition temperature, the copolymers formed colloidal nanoparticles with a dehydrated PNIPAAm core surrounded by DNA. When the formation of particles was conducted at higher temperatures, the dehydration of the copolymers proceeded such that the hydrodynamic radius (Rh) of the particles decreased. From the results of light scattering measurements, we calculated the surface area of particles occupied by one DNA (S(DNA)). The S(DNA) value decreased with increasing formation temperature, indicating that the DNA density on the particle surface increases with increasing formation temperature. The increase in the DNA density was also confirmed from the zeta-potential measurement of the particle. When MgCl2 was added to the copolymer solutions, the anionic charge of DNA was neutralized by Mg2+ so that Rh and the molecular weight of the particles increased with the increasing MgCl2 concentration. The turbidimetric detection of a target DNA was successfully demonstrated by utilizing the stability decrease of the colloidal particle upon hybridization on the particle surface.