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By using two oligonucleotides bearing a monophosphate group at the terminus and placing them at both edges of gap-site formed in substrate DNA, this site was preferentially and efficiently hydrolyzed by Ce(IV)/EDTA. In this study, site-selective hydrolysis of gap-site in long substrate DNA (274-mer) was investigated. When two monophosphate groups were placed to both edges of 5-base gap, predetermined site was selectively hydrolyzed by Ce(IV)/EDTA. Furthermore, desired recombinant DNA was prepared from this fragment and foreign DNA using T4 DNA ligase.

Manipulation of DNA through Ce(IV)/EDTA-induced site-selective hydrolysis using phosphate-bearing oligonucleotides