| Component | 50 μL | Final Concentration | |
|---|---|---|---|
| 1 | Nuclease–free water | 30.0 | |
| 2 | 5X PrimeSTAR GXL buffer | 10 | 1X |
| 3 | dNTP Mixture (2.5 mM each) | 4 | 200 μM |
| 4 | 10 μM sgRNA_forward | 2.5 | 0.5 μM |
| 5 | 10 μM sgRNA_reverse | 2.5 | 0.5 μM |
| 6 | PrimeSTAR GXL DNA Polymerase | 1.0 | 1.25 unit/50 μL reaction |
| total | 50 |
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