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Transfection mixture
| Component | Amount | Notes |
|---|---|---|
| Opti-MEM | 125 μL | Vortex 2–3 s. |
| lipo3000 | 5 μL | |
| Opti-MEM | 125 μL | Mix well, then add diluted DNA + P3000 reagent to diluted lipo3000. Incubate at 20°C–25°C for 15 min. Then add the 250 μL mixture to cells. |
| P3000 reagent | 5 μL | |
| pCMV-dR8.91 | 1 μg | |
| pMD2.G | 0.5 μg | |
| gRNA transfer vector | 1 μg |
