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Effects of CPT and arylstibonic acid on the DNA cleavage equilibrium. (A) hTopo forms a reversible phosphotyrosyl linkage with DNA. Upon trapping the covalent complex with SDS, a FAM-labeled 12-mer DNA strand is generated which is in direct proportion to the amount of covalent complex formed. (B) Separations of 22-mer DNA and 12-mer cleavage product by denaturing PAGE. The additions to each reaction are indicated. A DNA concentration of 10 nM was used.
