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. 2009 Aug 20;28(19):2981–2991. doi: 10.1038/emboj.2009.233

Figure 2.

Figure 2

Pol III HE releases DNA before it releases from β. (A) The reaction scheme to test effectiveness of a trap for free Pol III* is illustrated at the top. The trap is a 7.2-kb circular 3′dideoxy-terminated primed ssDNA, preloaded with a β clamp. Analysis of products in a native agarose gel at different amounts of added trap is shown to the left and quantification of DNA synthesis is shown in the plot to the right. (B) The experiment to test whether Pol III* stays bound to β after colliding with the downstream primer #2 is illustrated at the top. Pol III HE was assembled onto primer #1, and then the trap DNA (containing a preloaded clamp) was added along with primer #2 (as indicated). Replication was initiated and quenched at the indicated times followed by analysis in an alkaline agarose gel.