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| Step 1 thermal cycling conditions | ||
|---|---|---|
| Steps | Temperature | Time |
| 1 | 98 °C | 2 min |
| 2 | 8 °C | 1 min |
| 3 | Sound | *** Add reaction mix |
| 4 | 8 °C | 2 min |
| 5 | 16 °C (Ramp rate: 0.3 °C/sec) * | 1 min |
| 6 | 22 °C (Ramp rate: 0.3 °C/sec) * | 1 min |
| 7 | 28 °C (Ramp rate: 0.3 °C/sec) * | 1 min |
| 8 | 36 °C (Ramp rate: 0.3 °C/sec) * | 30 sec |
| 9 | 36.5 °C (Ramp rate: 0.3 °C/sec) * | 1 min |
| 10 | 37 °C | 8 min |
| 11 | Go to Step 1 | 3 times |
| 12 | 4 °C | Until removed from the cycler |
Add reaction mix in the 1st cycle but diluted Sequenase Enzyme in the remaining 3 cycles
