| Component | Amount/reaction (μl) |
| Linearized plasmid | 0.02–0.5 pmol (depends on DNA concentration) |
| Pre-sgRNA expression cassette (gBlocksTM) | 0.02–0.5 pmol (depends on DNA concentration) |
| Gibson assembly master mix (2 × ) | 10 |
| Nuclease-free water | Add up to 20 |
| Final volume | 20 |
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