(A) Illustration of the PME technology. Here, each laser operates in a CW mode with mechanical shutters pulsing the different excitation beams in sequential order. The single coaxial PME beam interrogates the fluorescently labeled DNA fragments, which are separated by capillary gel electrophoresis. Scattered laser light is rejected via specific long-pass or wavelength notch filters, with pulsed emission signals from the dye-labeled DNA fragments being detected by the photomultiplier tube (PMT) without use of any dispersing elements. (B) Unprocessed fluorescence data obtained during the electrophoretic run for the TCF1 exon 10 gene region using PME dye-primers. Blue, green, black, and red traces are AF-405, BODIPY-FL, 6-ROX, and Cy5.5 dye-primers terminated with ddCTP, ddATP, ddGTP, and ddTTP respectively. (C) Transformation of the raw trace data derived from the experiment described in B into readable, DNA sequence data using mobility software correction. Reprinted with permission from National Academy of Sciences, U.S.A. © 2005, Lewis et al. 2005.
