| Component | Amount/reaction (μl) |
| Plasmid (CasRx pre-sgRNA cloning backbone) | 2 μg (depends on DNA concentration) |
| NheI EcoRI | 1 (20 U) 1 (20 U) |
| CutSmart® buffer (10 × ) | 2 |
| Nuclease-free water | Add up to 20 |
| Final volume | 20 |
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