Assay Method Information

Assay Name:  RNA-dependent DNA Polymerase (RDDP) Assay
Description:  RNA-dependent DNA polymerase (RDDP) activity was measured as described[J. Microbiol. 2015, 53:288-293] in Tris HCl buffer (25 mL, 60 mm, pH 8.1) containing MgCl2 (8 mm), KCl (60 mm), DTT (13 mm), poly(A)-oligo(dT) (2.5 mm), dTTP (100 mm), and HIV-1 RT (6 ng, according to a dose-response curve). After enzyme addition, the reaction mixture was incubated for 30 min at 37 °C, then the reaction was stopped by addition of EDTA. Reaction products were detected by the addition of PicoGreen (code P7581 Thermo Fisher Scientific) according to the manufacturer's instructions and measured with the Victor 3 (PerkinElmer) equipped with filters (lex=502 nm, lem=523 nm).
Affinity data for this assay
 

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