Cleavage of DNA15-RNA1-DNA13/DNA29 substrate with Halo-RNase H2.

<p>The 5′-end labeled DNA15-RNA1-DNA13/DNA29 substrate was hydrolyzed by Halo-RNase H2 at 37°C for 15 min in the presence of 3 M NaCl and various concentrations of MnCl₂ or MgCl_2. The hydrolysates were separated on a 20% polyacrylamide gel containing 7 M urea. The concentration of the substrate was 1 µM. The amount of the enzyme added to the reaction mixture (10 µl) was 500 ng. Divalent metal ions used to hydrolyze the substrate and the concentrations of these divalent metal ions are shown above the gel. The arrows indicate the 5′-end labeled substrate (top) and 5′-end labeled 15-mer DNA (bottom).