| Reagent | Volume for 1x reaction (μl) | Incubation Time (hr) | Incubation Temperature (°C) |
| 450 ng of Gag-MJ4 plasmid | x μl for 450 ng | ||
| NEB CutSmart Buffer (previously NEB Buffer #4) | 2 | ||
| NgoMIV restriction enzyme | 0.5 | ||
| HpaI restriction enzyme | 0.5 | ||
| Nuclease-free H2O | x | ||
| Total volume | 20 | 2 | 37 |
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