|
| ||
| Component | Amount (μl) | Final concentration |
|
| ||
| Rapid Ligase Buffer, 2× | 12.5 | 1× |
| FastDigest Esp3I (BsmBI) | 1 | |
| DTT | 0.25 | 1 mM |
| BSA, 20 mg ml−1 | 0.125 | 0.1 mg ml−1 |
| T7 ligase | 0.125 | |
| Diluted oligo duplex from Step 72 | 1 | 0.04 μM |
| sgRNA library backbone | 1 | 1 ng μl−1 |
| UltraPure water | 9 | |
| Total | 25 | |
|
| ||
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