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2O to 10 μL. The samples were prepared with enzyme added last and incubated at 37 °C for 30-45 min. The reaction was stopped with 2.5 μL of decatenation buffer (5% sarkosyl, 25% glycerol, and 0.0025% bromophenol blue). The drugs were extracted from the incubation with 10 μL of 24:1 chloroform/isoamyl alcohol. The aqueous DNA containing samples were loaded on a 1% agarose gel
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2O. The DNA bands were detected on a UV light box and photographed with Polaroid 525 film. Controls were no enzyme, enzyme, m-AMSA (100 μM), and DMSO (1%).
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m-AMSA, the topo II-inhibitor utilized as a control in our DNA relaxation assay, is also a known DNA intercalator
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m-AMSA, the topo II-inhibitor utilized as a control in our DNA relaxation assay, is also a known DNA intercalator.
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