| Component | Amount | Final Concentration |
| Purified F1 PCR product (from step 46) | 3 μl | unknown |
| Purified F2 PCR product (from step 46) | 3 μl | unknown |
| Purified F3 PCR product (from step 46) | 3 μl | unknown |
| 10× Expand Buffer with MgCl2 | 5 μl | 1× |
| 10mM dNTPs | 4 μl | 0.8 mM |
| Nuclease-free water | 31.625 μl | |
| Expand Enzyme (3.5 U μl-1) | 0.375 μl | 0.02625 U μl-1 |
| Total | 50 μl | |