| Components for eukaryote method | Amount (μl) | Final |
|
| ||
| 5× Buffer | 10.0 | 1× |
| PLasmid from Step 15 (100 μg ml−1) | 0.25 | 25 μg |
| Primer 9 (5 mM) | 3.0 | 300 μM |
| Primer 10 (5 mM) | 3.0 | 300 μM |
| dNTP (2.5mM) | 5.0 | 250 μM |
| NucLease-free H2O | 25.25 | |
| DMSO | 2.5 | 5.0% (vol/vol) |
| Phusion hot start (2 U μl−1) | 1.0 | 2 U |
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