Table 2 Temperature profile for amplification and high resolution melting analysis of DQA1*05, DQB1*02 and DQB1*03:02 alleles
Amplification program | DQA1*05 | DQB1*02 and DQB1*03:02 | ||||
|---|---|---|---|---|---|---|
Temp | Time | Cycles | Temp | Time | Cycles | |
Hold | 95 °C | 5 min | 1 | 95 °C | 5 min | 1 |
Amplification 1 | 95 °C 65 °C 72 °C | 20 s 30 s 30 s | 20 | 95 °C 68 °C 72 °C | 20 s 30 s 30 s | 10 |
Amplification 2 | 95 °C 60 °C 72 °C | 20 s 30 s 30 s | 20 | 95 °C 63 °C 72 °C | 20 s 30 s 30 s | 25 |
HRM | 80–90 °C 0.1 °C increments for 2 s | 82–90 °C 0.1 °C increments for 2 s | ||||