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| | RNAse H-Polymerase-Independent Cleavage Assay |
| Description: | The RNase H activity associated with HIV-1 RT was measured in a 100 mL reactionvolume containing Tris HCl (pH 7.8, 50 mm), MgCl2 (6 mm), dithiothreitol (DTT, 1mm), KCl (80 mm), hybrid RNA:DNA (5'-GTTTT CTTTT CCCCC CTGAC-3'-fluorescein, 5'-CAAAA GAAAA GGGGG GACUG-3'-Dabcyl) and RT (3.8 nm). The reaction mixture was incubated for 1 h at 37 °C, the reaction was stopped by addition of EDTA, and products were measured with a multilabel counter plate reader (Victor 3, PerkinElmer) at λex=490 nm and λem=528 [[Esposito et al., Pathog. Dis., 68:116-124; Xu et al., J. Microbiol., 53:288-293]. |
| Affinity data for this assay | |
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