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. 2008 Aug 6;36(16):5281–5289. doi: 10.1093/nar/gkn507

Figure 5.

Figure 5.

TES reactions conducted with the C. albicans (rC) ribozyme. (A) Polyacrylamide gel of TES reactions using both 5′ and 3′-end radiolabeled substrates. Reactions were conducted using 75 nM rC, H0Mg (−) or H25Mg (+) buffer, for 30 min at 44°C. Lanes A, C and E contain 5′-end radiolabeled size controls, respectively. Lanes B, D and F contain 3′-end radiolabeled size controls, respectively. Note that the 3′-end radiolabeled size controls are one nucleotide larger than 5′-end radiolabeled size controls. Lanes G–J contain the normal rC ribozyme with 5′-end radiolabeled (lanes G and H) or 3′-end radiolabeled (lanes I and J) substrate. (B) Graphs of TES reactions using 5′-end radiolabeled substrate. All reactions were conducted as above except for the changing variable. (C) Graph of rGTP concentration dependence for TES reactions conducted with the rC ribozyme. All data points represent the average of at least two independent reactions, with standard deviations typically below 10%. Note for all graphs, the TES product is represented by filled triangles and the substrate cleavage product is represented as open triangles.