Table 2.
Construction of non-target DNA panels from initial High-Fidelity PCR amplifications of vouchered DNA samples
| Panel | H. androsaemum | H. kouytchense | H. maculatum | H. athoum | H. calycinum | H. ascyron | H. perforatum |
|---|---|---|---|---|---|---|---|
| Non-and |
x |
✓ |
✓ |
✓ |
✓ |
✓ |
✓ |
| Non-ath |
✓ |
✓ |
✓ |
x |
✓ |
✓ |
✓ |
| Non-asc |
✓ |
✓ |
✓ |
✓ |
✓ |
x |
✓ |
| Non-perf |
✓ |
✓ |
✓ |
✓ |
✓ |
✓ |
x |
| Multiplex | ✓ | ✓ | ✓ | ✓ | ✓ | ✓ | ✓ |
The initial PCR products were diluted by 1 × 10-5 (except H. athoum, 1 × 10-4) and equal volumes of each sample were added to the panel mixtures, as indicated by a tick. The panels are described by the name of the species that is not present, indicated by a cross. These panels were then used to test the species-specific primers for each target for cross-amplification, allowing six DNA samples to be tested simultaneously. The panel used to test the multiplex reaction contained all the available samples, and is named Multiplex.