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| Steps | Tube | Component | Per well of a 6-well plate |
|---|---|---|---|
| 1 | 1 | Opti-MEM™ I Medium | 100 µl |
| Lipofectamine™ Stem Reagent | 4 µl | ||
| 2 | 2 | Opti-MEM™ I Medium | 100 µl |
| DNA | 2 µg* | ||
| 3 | Add diluted DNA to diluted Lipofectamine Stem Reagent | ||
| 4 | Incubate for 10 min at room temperature | ||
| 5 | Add DNA-lipid complex to cells (200 µl/per well) | ||
| 6 | Incubate and monitor the transfected stem cells at 37 °C for 2 days | ||
*Equimolar amounts of Cas9 plasmid DNA and gRNA plasmid DNA were added
