|
| ||
| Component | Amount per reaction (μl) | Final concentration |
|
| ||
| NEBNext High Fidelity PCR Master Mix, 2× | 25 | 1× |
| Pooled sgRNA library template from Step 31 | 1 | 0.4 ng μl−1 |
| NGS-Lib-Fwd primer (unique) | 1.25 | 0.25 μM |
| NGS-Lib-KO-Rev or NGS-Lib-SAM-Rev primer (barcode) | 1.25 | 0.25 μM |
| UltraPure water | 21.5 | |
| Total | 50 | |
|
| ||
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