| Component | Amount | Final Concentration |
| Plasmid DNA | 5 μl (∼0.75 μg) | ∼25 ng /μl-1 |
| 10× EcoRI Buffer | 3 μl | 1× |
| EcoRI (20 U μl-1) | 1 μl | 0.66 U μl-1 |
| HindIII (20 U μl-1) | 1 μl | 0.66 U μl-1 |
| Nuclease-free water | 20 μl | |
| Total | 30 μl | |
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