Skip to main content
. Author manuscript; available in PMC: 2019 Mar 15.
Published in final edited form as: Methods. 2017 Dec 13;137:37–48. doi: 10.1016/j.ymeth.2017.12.007

Figure 6. Measuring ensemble FRET by RatioA method.

Figure 6

Spectra used for calculation of the ratioA in doubly-labeled EF-G•GTP (EF-G_301–541_Cy3/Cy5) [30]: (1) the doubly labeled sample emission excited at 540 nm (open circles); (2) the doubly labeled sample emission excited at 635 nm (triangles); (3) normalized emission of donor-only labeled sample excited at 540 nm (solid line); (4) the extracted acceptor fluorescence (black circles) obtained by subtracting (3) from (1). The ratioA is determined from (4) divided by (2).