FIG. 3.

Reverse transcription proceeds with protracted kinetics in highly purified resting T cells but can yield double-stranded HIV-1 DNA molecules of the appropriate length. Highly purified resting T cells were obtained from HIV-1-negative donors by cell sorting. Control MT-2 cells and resting CD4⁺ T cells were infected with HIV-1 IIIb at a multiplicity of infection of 1. At the indicated times, DNA was isolated and nested LM-PCR was performed to detect the U3 end of the linear HIV-1 genome. The LM-PCR involved an initial reaction of 15 cycles with outer LPCR-L and oligonucleotide 25t. Reaction products were diluted 1:4 in water, and a second 25-cycle PCR was performed using oligonucleotide 25t and LPCR-L. Reaction products were run on a 2% gel and Southern blotted with a U3-specific probe.