We unambiguously demonstrated DNA attraction and its regulation mediated by divalent cations Mg2+ and Ca2+ by tethering a DNA single chain at various pH solutions. It is found that DNA is compacted when the pH of the solution containing these divalent counterions is decreased below 5. When the pH of the medium is ∼4, DNA is in an unstable transition state, being able to switch between compact and extensible states. We can also regulate the DNA attraction through a cyclic process of DNA compaction and unraveling by alternating the pH of the solution between 3 and 8. The corresponding change of morphology of DNA modulated by pH is also confirmed by atomic force microscopy (AFM). In the theoretical aspect, the present experimental finding is consistent with the coarse-grained simulation of Langevin dynamics on the effect of pH on DNA in a solution of divalent counterions.