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| | DNA Polymerase Assay |
| Description: | The RNA-dependent DNA polymerase (RDDP) activity associated with HIV-1 RT was measured by use of the Invitrogen EnzCheck Reverse Transcriptase Assay Kit, as described [Costi et al., J. Med. Chem., 56:8588-8598], in a 50 mL volume containing Tris HCl (pH 8.1, 60 mm), MgCl2 (8 mm), KCl (60 mm), DTT (13 mm), dTTP (100 mm), HIV-1 RT (2 nm) and poly(A)-oligo(dT). The reaction mixture was incubated for 30 min at 37 °C. The enzymatic reaction was stopped by addition of EDTA, and products were measured with a Victor 3 (Perkin- Elmer) at 502/523 nm after picogreen addition [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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