Abstract This study applied TAM air isothermal calorimeter to measure the adsorption enthalpies of DNA on eight colloidal fractions from permanent-charge and variable-charge soils. The adsorption of DNA on soil colloids was also examined by equilibrium adsorption analysis. The data evaluated from isotherms fitted by Langmuirean model revealed that the affinity of DNA for variable-charge soil colloids was higher than that for permanent-charge soil colloids. More tightly bound DNA molecules were observed on coarse clays and inorganic clays than on fine clays and organic clays, respectively. The adsorption enthalpies of DNA on permanent-charge soil colloids were higher than those on variable-charge soil colloids. DNA adsorption on organic clays is endothermic, whereas that on inorganic clays is exothermic. Dehydration and electrostatic repulsion were considered to cause the higher adsorption enthalpies of DNA with organic clays, while hydrogen bonding, ligand exchange and electrostatic attraction result in the lower DNA adsorption enthalpies on inorganic clays. The thermodynamic parameters presented in this study have important implication for providing further insight into mechanisms of the adsorption of DNA on soil particles.