| Volume | Final concentration | |
|---|---|---|
| Multiplexed sgRNAs (from Step 24) | 5000 ng | 200 ng/μL |
| 20 μM EnGen Cas9 | 4.2 μL | 3.36 μM |
| 10x NEBuffer r3.1 | 2.5 μL | 1x |
| Nuclease free water | To 19 μL |
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