Masterclear cap strips

qPCR reactions were performed for all DNA extracts of the inclusivity/exclusivity panel for the established assay (Table 1) and run on a Mastercycler^® ep realplex S (Eppendorf, Hamburg, Germany). Each qPCR reaction was performed in a total volume of 25 μL, containing 5 μL template DNA, 8.7 μL sterile ddH₂O, 1.3 μL primer-probe-mix (primers: 18 μM and probe: 5 μM in the stock), and 10 μL TaqMan Environmental Master Mix (Applied Biosystems, Foster City, California) avoiding PCR inhibition [27 (link)]. Eppendorf real-time PCR tube strips covered with Masterclear^® cap strips (Eppendorf, Hamburg, Germany) were used as reaction vessels for amplification and detection. The qPCR temperature profile entailed the following steps: initial denaturation (2 min at 50°C) for optimal Uracil-N-Glycosylase enzyme activity and successive activation of the DNA polymerase (10 min at 95°C), followed by 50 cycles of 15 sec at 95°C and 1 min at 60°C. Two negative controls (environmental control and extraction blank control) were included at least once in qPCR procedures to exclude any contamination.