Fluorescence Signal from Packaging Complexes

<p>The graphs show the fluorescence intensity of vertical (black) and horizontal (red) polarization detected simultaneously. One example for each of the six investigated mutants is shown. The data were recorded with an integration time of 75 ms and three-points smoothing was applied. The camera background (closed shutter) was subtracted. The signal was normalized using the fluorescence intensity of the bead after packaging was completed. The traces are vertically shifted for clarity. The inset shows a zoom of the data. Here, the unfiltered data is displayed (scatter) together with the three-point sliding average. (A) Fluorescence recorded for a single complex of mutant 170C. The shutter is opened after about half a second. After t = 4 s the first dye bleaches and after 19 s the second. After about t = 100 s the fluorescence signal again starts to increase, since the magnetic bead is pulled into the evanescent field as the prohead reels in the DNA. The signal increases in an exponential fashion as the fluorescent bead samples the intensity profile of the evanescent wave. Similar behavior was observed for (B) 97C; (C) 168C; (D) 189C; (E) 190C; and (F) 260C.