| Component | Volume (μL) | Final concentration |
|---|---|---|
| 2× Q5 Master Mix | 15 | 1× |
| 10 μM locus-specific oligo | 1.5 | 0.5 μM |
| 10 μM universal oligo | 1.5 | 0.5 μM |
| Plasmid with positive/negative selection cassette (10 ng/μL) | 1.0 | 0.33 ng/μL |
| Nuclease-free water | 11 | — |
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