Table 3. Glycoprotein B gene PCR amplification conditions.
| Glycoprotein B amplification conditions | |||
|---|---|---|---|
| gB amplification | Primer sets | Amplicon size (nt) | Cycling conditions |
| Full (from fresh tissues) | gBTeHV3FW-gBTeHV3RV | 2,484* | 95°C for 1’ + 40 cycles = 95°C for 30”, annealing at 50°C for 30” + 68°C for 1’. Final elongation step at 68°C for 10’ |
| Full (from formalin-fixed tissues) | FWA-RVA | 524* | 95°C for 15’ (hot start)+ 45 |
| FW1-RVB | 409* | cycles = 95°C-1’ + annealing | |
| FW2-RVC | 413* | at 50°C (FWA and RVA, | |
| FW3-RVD | 409* | FW4 and RVE, FW5 and | |
| FW4-RVE | 405* | RVF) or 52°C (FW1 and | |
| FW5-RVF | 389* | RVB, FW2 and RVC, FW3 | |
| FW6-gBTeHV3RV | 387* | and RVD, FW6 and | |
| Partial | FW4-RVE | 405* | gBTeHV3RV) for 30”, + |
| FW5-RVF | 389* | 72°C for 30”. A final | |
| FW6-gBTeHV3RV | 387* | elongation step for 10’ at 72°C | |
* Expected amplicon size including primers