We identified a new class of deoxyribozymes named A-2 and A-3 by in vitro selection which required both Cu2+ and Mn2+ as cofactors that selectively and rapidly cleave the DNA substrate. Studies confirmed that they cleaved via a mechanistic pathway involving the formation of hydrogen peroxide as the reactive species. The kinetics, secondary structures and sequence tolerance of the new class of the deoxyribozymes A-2 and A-3 were reported.