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PCR protocol for genotyping using advanced ASQ method for SNP genotyping.
| Step | Temperature | Duration (sec) | Notes |
|---|---|---|---|
| 1 | 95°C | 120 | Initial denaturation |
| 2 | 95°C | 10 | First-round denaturation |
| 3 | 55°C | 20 | First-round annealing |
| 4 | 68°C | 20 | First-round extension |
| 5 | 10 cycles repeat for steps 2–4 | ||
| 6 | 95°C | 10 | Second-round denaturation |
| 7 | 68C | 30 | Second-round annealing and extension |
| 8 | 55°C | 30 | FRET cassette annealing and signal read |
| 9 | 30 cycles repeat for steps 6–8 |
