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Procedure
DNA substrate (10nM) containing 8-oxodGTP:rA was incubated with 0, 0.5, 1.5, 2.5 and 5Units of purified RNase H2 protein in buffer containing 20Mm Tris-HCl, 2mM MgCl2, 10mM KCl, 10mM (NH4) SO4, and 0.1% Triton X-100. The reaction was incubated at 37°C for 1h. The sample were separated by 20% denaturing PAGE and fluorescent band detected by Typhoon scanner (GE Healthcare).
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Cilli Piera
Ph.D. | Pescara