Table 1 Conditions of multiplex PCR
From: Comparison of blood glucose levels and allergic responses on treatment with six wheat cultivars
Multiplex PCR | Glu-1 | Glu-A3 | Glu-B3 |
|---|---|---|---|
Allele | Glu-Alc/Glu-Blbcf/Glu-Dld | Glu-A3ac/Glu-A3d/Glu-A3e | Glu-B3d/Glu-B3 fg/Glu-B3 h |
PCR reaction components | |||
Taq DNA polymerase (U) | 2 | 2 | 2 |
Each dNTP (mmol/l) | 2 | 2 | 2 |
One gene (μmol/l) | |||
Concentration of DNA (ng) | 100 | 100 | 100 |
PCR amplification conditions | |||
Pre-denature | 94 °C, 15 min | 94 °C, 5 min | 94 °C, 8 min |
Denature | 94 °C, 40 s | 94 °C, 1 min | 94 °C, 1 min |
Annealing | 60 °C, 1 min | 60 °C, 1 min | 58.5 °C, 1 min |
Extension | 72 °C, 1 min | 72 °C, 1 min | 72 °C, 2 min |
No. of cycles | 30 | 35 | 40 |
Final extension | 72 °C, 5 min | 72 °C, 5 min | 72 °C, 10 min |
Gel electrophoresis | |||
Agarose (%) | 1.0 | 1.0 | 1.0 |
Voltage (V) | 100 | 100 | 100 |
Time (min) | 40 | 40 | 40 |