Novel 3′-Processing Integrase Activity Assay by Real-Time PCR for Screening and Identification of HIV-1 Integrase Inhibitors

Figure 4

Integrase activity in conventional PCR. Biotinylated HIV-1 LTR substrates were added in reaction mixture (lanes 1–12) to a final concentration at 10 pM (lanes 1, 4, 7, and 10), 1 pM (lanes 2, 5, 8, and 11), and 0.2 pM (lanes 3, 6, 9, and 12), respectively. Lanes 1–3: the reaction mixture without IN; lanes 4–6: the reaction mixture with IN; lanes 7–9: the reaction mixture with IN and Raltegravir to a final concentration at 10 μM; lanes 10–12: the reaction mixture with IN and NaN₃ to a final concentration at 5%; lanes 13-14: IN and reaction buffer only as negative control.