| Reagents | Volume (μl) | Final |
|---|---|---|
| 5x Phusion HF buffer | 10 | 1x |
| dNTPs (2.5 mM each) | 4 | 200 μM |
| P5-I5 (10 μM) | 2 | 400 nM |
| P7-I7 (10 μM) | 2 | 400 nM |
| Phusion polymerase (2 U/μl) | 0.5 | 1 U |
| DNA products (from step 39 or step 35 if blocking is omitted) | 15 | - |
| H2O | 16.5 | - |
| Total | 50 |
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