Manipulation of DNA presents a great interest in biotechnology and therapeutics. The molecules that damage DNA selectively offer new prospects for controlled manipulation of DNA. The conjugations of DNA-code reading molecules such as polyamides to reagents that induce DNA damages provide an approach to reach this goal. In this work, a new compound which contained polyamide and ascorbic acid conjugated by flexible linker (polyamide-Vc), was successfully synthesized, characterized, and evaluated as DNA cleavage agent, compared with that by using ascorbic acid molecule. The kinetics data showed that polyamide-Vc successfully promoted the cleavage of plasmid DNA, with k(max) of 0.314 h(-1) and K(d) of 0.105 mM. The evaluation of DNA linearization elicited that the activity of cleaving double-strand in the supercoiled pUC18 plasmid DNA by polyamide-Vc was enhanced remarkably, achieving n1/n2 ratio of 13.9 at 1.2 mM for 1 h. The introduction of polyamide to Vc could also partially weaken the inhibition of hydrogen radical to double-strand cleavage process because of its good binding activity to DNA. We anticipate that this work could provide a method for improving the efficiency of double-strand cleavage, especially to oxidative cleavage agents.