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| Reagent | Amount |
| gRNA cassette plasmid with connectors | 20 fmol |
| Cen6-Ura cassette* | 20 fmol |
| Cas9 plasmid (pYTK036) | 20 fmol |
| NEB T4 buffer 10x | 1.0 µl |
| NEB T7 ligase | 0.5 µl |
| NEB BsmBI | 0.5 µl |
| ddH2O | To 10 µl |
