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Role of Topo I. The 69 bp duplex after preincubation with 2 µM 16TCG in the described buffer was treated with different Topo I mutants: lane 1, no enzyme; lane 2, Topo I mutant Y723F; lane 3, Topo I wild-type; lane 4, Topo I mutant ΔC; lane 5, Topo I mutant ΔN. After protease digestion, the samples were loaded on a sequencing gel. The presence of the band marked with an asterisk was dependent on the Topo I preparation in the experiment and was not further characterised.
