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Figure 3 | Retrovirology

Figure 3

From: Detection, characterization and regulation of antisense transcripts in HIV-1

Figure 3

Specific detection of HIV-1 antisense transcript in infected cells lines and transfected 293T cells. Synthesis of cDNA was performed on polyA+ RNA using 24-6 (control RT primer) or 24-6F (floating RT primer) and purified on a PCR-cleanup column. PCR amplifications were performed using the reverse primer 30-20 (anchor) and forward primers 26-5 (lane 1, 3, 5) or 25-3 (lane 2,4 and 6) in order to specifically amplify 24-6F-synthesized cDNA. Samples were tested for anchor primer specificity (lanes 1–2) and cDNA cleanup efficiency (lanes 3–4). Lane 5 and 6 show specific amplification of HIV-1 antisense transcript from J1.1 OM10.1, U937 and ACH-2 chronically infected cells lines and 293T transfected with complete pNL4.3 proviral DNA or 5'LTR-deleted pNL4ΔNar1 construct.

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