| Order | Temperature (°C) | Duration (min) | Purpose |
|---|---|---|---|
| 1 | 30 | 25 | Filling in cut MspI sites |
| 2 | 37 | 25 | A-tailing |
| 3 | 70 | 10 | Inactivation of Klenow exo- |
| 4 | 4 | Hold | — |
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