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. 2013 Mar 5;8(3):e58177. doi: 10.1371/journal.pone.0058177

Figure 2. Workflow of pre-amplification experiments.

Figure 2

A fresh aliquot of template DNA (linearised ADH plasmid or Arabidopsis gDNA) was diluted in carrier to a ‘high’ concentration (3000 copies/µl to give λ = 0.76) and a ‘low’ concentration (500 copies/µl to give λ = 0.13). The ‘low’ concentration was used as the template in the pre-amplification reaction. The pre-amplification reaction was serially diluted in 1× TE (pH 8.0) and using qPCR, the dilution to give an approximately λ = 0.76 for the Adhβ target was selected. dPCR analysis using the 48.770 arrays was performed with the Adhα-FAM:Adhβ-VIC duplex assay for the ‘high’ and ‘low’ concentration non-amplified template DNA and diluted pre-amplified template DNA. This experimental workflow was repeated on three separate days. A further three experiments were performed on three separate days using the Adhδ-FAM:Adhβ-VIC and Adhδ-VIC:Adhβ-VIC duplex assays.