A | B | C | |
Component | Amount in ul -in each 0.2 mL tube | Negative Control tube | |
PCR water | variable up to 20ul | variable up to 20ul | |
Thermopol buffer 10x | 2 | 2 | |
dNTP Mix 10 mM | 0.4 | 0.4 | |
Fw/Rv primers | 1/1 | 1/1 | |
DNA template | 0.5 (depending on the concentration) | / | |
Enzyme | 2 units (1ul) | 2 units (1ul) |
A | B | C | D | |
Phase cycle | Temperature (°C) | Duration (sec) | Number of cycles | |
Initial denaturation | 95 | 120 | ||
Denaturation | 95 | 30 | ||
Annealing | 50 to 65 (depending on primers) | 20 | 30 to 35 | |
Extension | 72 | variable based on primers | ||
Final extension | 72 | 120 |