| Component | Volume | Final Concentration |
| 10 X NEBufferTM r3.1 (NEB# B6003) | 3 μL | 1 X |
| 300 nM gRNA | 3 μL | 30 nM |
| 1 μM SpCas9-His | 1 μL | 30 nM |
| Nuclease free water | 20 μL |
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