Fig. 1

qBiCo-v1 assay development using synthetic DNA standards. (A) Amplification curves of the single qPCR assays for the eight serially diluted DNA standards, each labelled with a different fluorescent dye; (B) Amplification curves of an example 5-plex qBiCo assay (STD3, in duplicate); (C) Performance of negative controls during 16 qPCR runs to determine the threshold where no false positive signals are obtained; (D) Standard curves of the four assays (50 − 0,39 ng/µl) used to calculate the concentration of each fragment in a sample; (E) Heatmap of the detection limit of each individual assay based on the serially diluted DNA standards; (F) Spike-in control performance in the serially diluted DNA standards during six qPCR runs; (G) Assay efficiency using the same DNA standard dilution series within five days. RFU: Relative fluorescent units; STD: Synthetic standard; IPC: Internal positive control; Cq: Quantification cycle