| Component | Volume per reaction (µl) | Final concentration |
|---|---|---|
| Pooled PCR 1 from step 12| | 10 | 1 nM |
| Phusion HF buffer (5x) | 20 | 1x |
| Phusion MgCl2 Solution (50 mM) | 4 | 2 mM |
| dNTPs (5 mM) | 5 | 250 µM |
| Primer IlluminaPCR2a (10 µM) | 6 | 0.6 µM |
| Primer IlluminaPCR2b (10 µM) | 6 | 0.6 µM |
| Phusion (2 U·µl-1) | 0. 5 | 1.0 U per 100 µl |
| Water | up to 100 | - |
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