"above/on/below": number of real-simulated comparisons above, on and below the diagonal, with "above" indicating single copy concentration.
Ideally, all probes should be sensitive enough to detect DNA at single-copy concentrations.
The threshold cycle (CT) values determined were plotted against the logarithm of their initial copy concentrations.
However, the relative standard deviation from the mean total 16S rRNA gene copies per concentration tested over time (n = 12, rsd = 60% for the highest, and rsd = 50% for the lowest concentration tested) did not exceed the relative standard deviation observed from the mean total 16S rRNA gene copies detected after 24 h (mean 16S rRNA gene copies for all PCB 61 concentrations, n = 21, rsd = 73%).
As the number of reactions increases, the confidence associated with the predicted copy number and the copy number concentration improves dramatically.
Since the calculation of the copy number concentration includes dividing the copy number estimate by the assay volume, knowing the correct partition or droplet volume is a key factor when measuring DNA concentrations using digital PCR [ 29].
Thus, the average value of the two measurement results was used as the reference value for the plasmid copy number concentration, which was (5.19 ± 0.22) × 10 copies μL−1.
The number of target DNA molecules of dilution S7 in ddPCR replicates is the most variable, because the copy number concentration of dilution S7 is the lowest (2.3 copies 20 μL−1).
The average copy number concentration of the three subsamples was 5.05 × 10 copies μL−1, with a relative standard deviation of 3.63 %.
> -wrap-foot> Cthe the average Ct value for the highest copy number/DNA concentration, Ct B the average Ct value for the lowest copy number/DNA concentration aThe DNA was diluted four times in a 1 2 series.
The large number of reactions in this ddPCR system allowed very precise copy number estimates which resulted in a relative expanded uncertainty of less than 5% for Lambda DNA copy number concentration.
