| Component | Amount (µL) |
| PBS | 1 |
| crRNA:tracrRNA duplex (from step 4.2) | 2 (200 pmol) |
| Alt-R Cas9 endonuclease (61 µM stock) | 2 |
| Total volume | 5 ul |
| Component | Amount (µL) |
| PBS | 1 |
| crRNA:tracrRNA duplex (from step 4.2) | 2 (200 pmol) |
| Alt-R Cas9 endonuclease (61 µM stock) | 2 |
| Total volume | 5 ul |