Table 2 The Fluidigm ‘AA 48 × 48 Standard v1’ PCR thermal protocol[8]
PCR stages | Temperature | Number of Cycles |
|---|---|---|
Thermal mixing and hot start phase | 50°C 2 minutes |  |
70°C 20 minutes | 1 | |
95°C 10 minutes |  | |
|  | 95°C 15 seconds |  |
PCR cycle | 60°C 30 seconds | 10 |
|  | 72°C 1 minute |  |
C0t cycle | 95°C 15 seconds | 2 |
80°C 30 seconds | ||
60°C 30 seconds | ||
PCR cycle | 72°C 1 minute | 8 |
95°C 15 seconds | ||
60°C 30 seconds | ||
72°C 1 minute | ||
C0t cycle | 95°C 15 seconds | 2 |
80°C 30 seconds | ||
60°C 30 seconds | ||
72°C 1 minute | ||
PCR cycle | 95°C 15 seconds | 8 |
60°C 30 seconds | ||
72°C 1 minute | ||
C0t cycle | 95°C 15 seconds | 5 |
80°C 30 seconds | ||
60°C 30 seconds | ||
Extension | 72°C 1 minute | 1 |