TABLE 2.

PCR and RT-PCR cycle conditions

Target	Conditions for:
	RT	Denaturation	PCR cycles
Total HIV-1 DNA	—a	95°C, 8 min	95°C, 10 s, 60°C, 10 s, 72°C, 6 s for 50 cycles
2-LTR circle	—	95°C, 8 min	95°C, 10 s, 66°C, 10 s, 72°C, 10 s for 15 cycles, then 35 cycles with the annealing temp decreased by 0.5°C/cycle to 59°C
Integrated HIV-1 DNA (first-round PCR)	—	95°C, 8 min	95°C, 10 s, 60°C, 10 s, 72°C, 170 s for 12 cycles
Integrated HIV-1 DNA (second-round PCR)	—	95°C, 8 min	95°C, 10 s, 60°C, 10 s, 72°C, 9 s for 50 cycles
Unspliced HIV-1 RNA	61°C, 25 min	95°C, 2 min	95°C, 10 s, 60°C, 40 s for 50 cycles
Multiply spliced HIV-1 RNA	61°C, 25 min	95°C, 4 min	95°C, 5 s, 54°C, 10 s, 72°C, 8 s for 50 cycles
U5-U3 HIV-1 RNA	61°C, 25 min	95°C, 2 min	95°C, 5 s, 60°C, 10 s, 72°C, 10 s for 50 cycles
CD3ɛ RNA	61°C, 20 min	95°C, 2 min	95°C, 15 s, 60°C, 40 s for 50 cycles
Cyclophilin A RNA	61°C, 30 min	95°C, 2 min	95°C, 1 s, 56°C, 10 s, 72°C, 15 s for 50 cycles

^a—, not applicable.