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| Reagent | Volume |
|---|---|
| 5× Phusion HF buffer | 10 μL |
| dNTPs (10 mM) | 1 μL |
| Ligation forward primer | 2 μL |
| Ligation reverse primer | 2 μL |
| pISceI-XsgRNA-2 (0.1 ng/μL) | 1 μL |
| Phusion high-fidelity DNA polymerase | 0.5 μL |
| H2O | To 50 μL |
