Figure 2.

Mutational analysis of mini-TAR RNA–DNA annealing in the absence of NC. Annealing was performed as described in ‘Materials and Methods’ section. (A) Each sample was divided in two aliquots which were analyzed by 4% agarose gel electrophoresis at 4°C in TBM buffer and at 25°C in TBE buffer. Nucleic acids were visualized by ethidium bromide staining (top) and autoradiography (bottom). Lane 1, heat-denatured mini-TAR ³²P-RNA. Mini-TAR ³²P-RNA was incubated in the absence (lane 2) or presence of unlabeled mini-cTAR DNAs (lanes 3–6). Mini-cTAR ³²P-DNAs were incubated in the absence (lanes 10–12) or in the presence of unlabeled TAR RNA (lanes 7–9). Monomeric and dimeric forms of mini-TAR RNA are indicated by m1 and d1, respectively. Monomeric and dimeric forms of mini-cTAR DNAs are indicated by m2 and d2, respectively. Loose and tight heteroduplexes are indicated by h1 and h2, respectively. (B) The table summarizes the results of the mutational analysis.