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FIGURE 6.

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PPi- and ATP-dependent unblocking of EFdA-MP-terminated primers. A, purified Td31/Pd18-EFdA-MP or Td26/Pd18+5-EFdA-MP-dTMP was incubated with HIV-1 RT in the presence of 6 mm MgCl2 and 150 μm PPi at 37 °C. Aliquots were removed, and reactions were stopped at the indicated time points (0–30 min). The excision products are shown in braces. B, purified Td31/Pd18-EFdA-MP or Td26/Pd18+5-EFdA-MP-dTMP was incubated at 37 °C with HIV-1 RT in the presence of 10 mm MgCl2, 3.5 mm ATP, and d(d)NTPs (100 μm dATP, 0.5 μm dTTP, and 10 μm ddGTP for Td31/Pd18-EFdA-MP or 100 μm dTTP and 10 μm ddCTP for Td26/Pd18+5-EFdA-MP-dTMP). Aliquots of the reactions were stopped at the indicated time points (0–90 min). C, purified Td31(6X)/Pd18+5-EFdA-MP (where X = T, A, C, or G) was incubated with HIV-1 RT in the presence of 6 mm MgCl2 and 150 μm PPi at 37 °C. Aliquots were removed, and reactions were stopped at the indicated time points (0–20 min). The excision products are shown in braces. The lane numbers are shown below each gel.

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