| Quantitative RT–PCR conditions | |
|---|---|
| Holding stage | Step 1: 95°C 20 s |
| Cycling stage | Step 2: 95°C 3 s |
| Step 3: 60°C 30 s | |
| Return to step 2: 40 times | |
| Melt curve stage | Step 4: 95°C 15 s |
| Step 5: 60°C 60 s | |
| Step 6: 95°C 15 s | |
| Step 7: 60°C 15 s | |
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