| Component | Amount/reaction (μl) |
| Plasmid (CasRx pre-sgRNA cloning backbone) | 2 μg (depends on DNA concentration) |
| BbsI | 1 (20 U) |
| CutSmart® buffer (10 × ) | 3 |
| Nuclease-free water | Add up to 30 |
| Final volume | 30 |
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