Fig. (2) Neither NS5B WT nor NS5B D8 mutant is affected by NS5A in the synthesis of the first 3 nt during replication in vitro. Either NS5B WT (0.9µM) or NS5B D8 (0.2 µM) NS5B was incubated with NS5A (2.6 pM, 26 pM, 260 pM, 2.6 nM, 26 nM, 260 nM) for 30 min on ice. Then the nucleotides (10 µM ATP, 3mM GTP, and 10 µCi [³²P] ATP) were added along with the LTMOA template (0.5 µM) and incubated for 30 min at 30°C. The products of the reaction were separated on a 23% PAGE-7M Urea gel.