| Reagent | Volume for 1x reaction (μl) |
| Nuclease-free H2O | 35.5 |
| 5x Phusion HF Buffer | 10 |
| dNTPs (40 mM deoxynucleotides) | 1 |
| Forward primer MJ4For1b (20 μM concentration) | 1 |
| Reverse primer MJ4Rev (20 μM concentration) | 1 |
| Phusion Hot Start II Polymerase | 0.5 |
| MJ4 plasmid as template (10 ng/ul) | 1 |
| Total volume | 50 |
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