| Component | Amount (μL) | Final concentration |
|---|---|---|
| Phusion U Green Multiplex Master Mix, 2× | 12.5 μl | 1× |
| PCR2 forward primer, 10 μM | 1.25 μl | 0.5 μM |
| PCR2 reverse primer, 10 μM | 1.25 μl | 0.5 μM |
| PCR1 unpurified product (Step 30) | 1 μl | - |
| Nuclease-free H20 | 9 μl | - |
| Total reaction volume | 25 μl | - |
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