FIG. 6.
HMA for the V3-V5 region of Env. The V3-V5 regions of Env for HIV-1 isolates were PCR amplified as outlined in Materials and Methods. Ten microliters of PCR product were used for homoduplex formation, while equal amounts (5 μl) of PCR products derived from different HIV-1 virus DNA were mixed to form heteroduplexes. The PCR products were denatured for 2 min at 94°C followed by rapid cooling in ice water. The samples were resolved on a nondenaturing polyacrylamide gel (6%) at a constant voltage of 250 V for 2.5 h. The gel was stained in ethidium bromide solution and visualized under UV light. Lanes: M, 1-kb molecular weight markers; 1, HIV-1JC16 homoduplex; 2, HIV-1JC16-HIV-1JC (genomic DNA) heteroduplex; 3, HIV-1JC (genomic DNA) homoduplex; 4, HIV-1JC16-HIV-1SF162 heteroduplex; 5, HIV-1JC16-HIV-1ZM18 heteroduplex; 6, HIV-1NC7 homoduplex; 7, HIV-1NC7-HIV-1NC (genomic DNA) heteroduplex; 8, HIV-1NC (genomic DNA) homoduplex; 9, HIV-1JC16-HIV-1NC7 heteroduplex; 10, HIV-1NC7-HIV-1SF162 heteroduplex; 11, HIV-1NC7-HIV-1ZM18 heteroduplex. HIV-1SF162 (HIV-1 subtype B3) and HIV-1ZM18 (subtype C2) plasmid DNAs are provided in the HMA kit as control DNAs. ssDNA, single-stranded DNA.