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

Figure 4.

Figure 4.

TES reactions conducted with the T. thermophila (rT-X) ribozyme. (A) Polyacrylamide gel of TES reactions using either 5′ or 3′-end radiolabeled substrates. Reactions were conducted using 166 nM rT-X, H0Mg (−) or H10Mg (+) buffer, for 45 min at 44°C. Lanes A, C and F contain 5′-end radiolabeled size controls, respectively. Lanes B, D and E 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 rT-X 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 rT-X 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.