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. 2005 Nov;11(11):1678–1687. doi: 10.1261/rna.2125305

FIGURE 3.

FIGURE 3.

Time course of Azoarcus-ribozyme-directed recombination of the B16–19RC ribozyme (152 nt) from the constituent oligonucleotides SL-5 and SL-6. In all lanes, only the SL-6 RNA oligonucleotide (Table 1) initially has 5′ radiolabel (*). (Lane 1) Unincubated *SL-6. (Lane 2) Negative recombination control in the absence of SL-5: 0.2 μM *SL-6 and 1.5 μM Azoarcus ribozyme incubated for 30 min. (Lane 3) Negative recombination control in the absence of Azoarcus ribozyme: 0.2 μM *SL-6 and 0.2 μM SL-5 incubated for 30 min. (Lanes 4–14) Recombination time course: 0.2 μM *SL-6, 0.2 μM SL-5, and 1.5 μM Azoarcus ribozyme. At 90 min, the recombination frequency (r.f.) has reached 17%. (Lane 15) 5′-end-labeled B16–19 RNA as a 152-nt size control. All reactions performed at 60°C in 10 mM MgCl2, 5 mM CaCl2, and 30 mM EPPS, nominal pH 7.6.