| Reagent | Volume (μl) | Final |
|---|---|---|
| DNA template | 5 | |
| Forward primer, 25 μM | 1 | 0.2 μM |
| Reverse primer, 25 μM | 1 | 0.2 μM |
| dNTP, 10 mM | 2 | 0.2 mM |
| pfuTurbo C× buffer, 10× | 5 | 1× |
| pfuTurbo C× polymerase (2.5 U μl−1) | 1 | 0.05 U |
| ddH2O | Up to 50 |
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