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Fig. 3 Multiple target detection (genes of interest) using EvaGreen-based ddPCR. (a) One-dimensional droplet
plots of resulting duplex assay optimized with different primer concentration. The different target genes show
different amplitudes. (b) Two-dimensional droplet plots of duplex assay. In all cases, manual thresholds were
drawn to assign clusters. The different target genes form different clusters
Fig. 4 Reaction preparation containing only one primer set at each well. They are used to identify the amplifica-
tion cluster of each gene. Well one (left column) contains both primer sets for the duplex detection; well two
(middle column) and well three (right column) contain two different primer sets to enable identification of the
different clusters within the first well. In all cases, manual thresholds are drawn to assign clusters
Katherine T. W. Lee et al.