| Reagents | Volume (μl) | Final |
|---|---|---|
| 5x Phusion HF buffer | 10 | 1x |
| dNTPs (2.5 mM each) | 1.5 | 75 μM |
| Bio-primer (1 μM) | 0.5 | 10 nM |
| Phusion polymerase (2 U/μl) | 0.5 | 1 U |
| sonicated DNA (from step 8) | 25 | 1-10 μg |
| H2O | 12.5 | - |
| Total | 50 |
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