PCR Cycling Conditions.
| Steps | Temperature | Time | Cycles |
|---|---|---|---|
| Initial Denaturation | 98 °C | 3 min | 1 |
| Denaturation | 98 °C | 20 sec | 30–35 cycles |
| Annealing | 60–62 °C | 20 sec | |
| Extension | 72 °C | 30 sec | |
| Final extension | 72 °C | 7 min | 1 |
| Hold | 4 °C | forever | |
| Steps | Temperature | Time | Cycles |
|---|---|---|---|
| Initial Denaturation | 98 °C | 3 min | 1 |
| Denaturation | 98 °C | 20 sec | 30–35 cycles |
| Annealing | 60–62 °C | 20 sec | |
| Extension | 72 °C | 30 sec | |
| Final extension | 72 °C | 7 min | 1 |
| Hold | 4 °C | forever | |