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Thermal cyclic stages of the qPCR assay.
| Thermal cyclic stage | Steps | Temperature (°C) |
Duration |
|---|---|---|---|
| Hold stage | Step 1 | 50.0 | 2 min |
| Step 2 | 94.0 | 5 min | |
| PCR stage (35 cycles) | Step 1 | 94.0 | 30 sec |
| Step 2 | 66.0 | 1 min | |
| Melt curve stage | Step 1 | 72.0 | 50 sec |
| Step 2 | 66.0 | 1 min | |
| Step 3(dissociation) | 95.0 | 1 sec |
