“…Total cellular RNA was extracted using RNAzol (Bioprobe Systems, Montreuil, France) and was amplified using rTth Polymerase (Perkin-Elmer, Norwalk, Conn.) in the presence of the GAG06-GAG04 primer pair to detect the HIV-1 unspliced Gag-Pol mRNA and the BSS-KPNA primer pair to detect the singly spliced mRNA, as described elsewhere (2,17,41,57). To measure proviral DNA levels, total cellular DNA was extracted and subjected to 20 or 40 repeated rounds of amplification with Amplitaq Gold DNA polymerase (Perkin-Elmer).…”