Fig. 5.

Compositional dynamics of fluorescent ligands on the custom RS. (A) Schematic of the expected signal sequence and example trace of ribosome compositional dynamics during late initiation showing that, although there is bleedthrough between Cy3 and Cy3.5 channels as well as Cy5 and Cy5.5 channels, the signal is sufficient to distinguish between the spectral channels. (B) Lifetimes for the Cy3-30S, Cy3.5-50S, and Cy5-Phe. Number of molecules analyzed was n = 307. Only a portion (∼5%) of the entire SMRT Cell was analyzed. (C) Arrival times for the Cy3.5-50S and Cy5-Phe tRNA. All error bars are SEMs. (D) Postsynchronization of the arrival of Cy5-Phe tRNA to the arrival of the Cy3.5-50S shown as a heatmap. The results were comparable to what was observed previously (25).