Allelic discrimination assays for the two subspecies of A. bicolor . The two fluorescent reporter dyes of VIC and FAM were used for subspecies-specific probes of A. bicolor bicolor and A. bicolor pacifica, respectively (Figure 1), and fluorescence intensities of each dye are plotted. The no template control (NTC) is shown with a cross symbol. (a) Various amounts of template DNA were tested for both A. bicolor bicolor (open circles) and A. bicolor pacifica (closed circles): a, 50 ng; b, 5 ng; c, 500 pg; d, 50 pg; e, 5 pg; f, 500 fg; g, 50 fg in each reaction. (b) Three and four haplotypes in A. bicolor bicolor (open circles; BBI-HT1-3) and A. bicolor pacifica (closed circles; BPA-HT1-4), respectively, were tested (Figure 2). (c) The ten anguillid species were tested with the two subspecies of A. bicolor, and the first three characters of each species name are shown instead of symbols. beb, A. bengalensis bengalensis; bel, A. bengalensis labiata; mos, A. mossambica; cel, A. celebesensis; jap, A. japonica; luz, A. luzonensis, bor, A. borneensis; int, A. interioris; mar, A. marmorata; obs, A. obscura.