A | B | C | |
Component | Volume | Final conc. | |
Template (< 1 µg) | X µL | - | |
Forward Primer (10 µM) | 1 µL | 0.4 µM | |
Reverse Primer (10 µM) | 1 µL | 0.4 µM | |
2x Master Mix | 12.5 µL | - | |
ddH20 | up to 25 µL | - | |
Total volume | 25 µL | - |
A | B | C | D | |
Step | Temp | Sec | Cycle | |
Initial denaturation | 95 ºC | 180 | ||
Denaturation | 95 ºC | 30 | 35 cycles | |
Annealing | 53-58 ºC | 30 | ||
Extension | 72 ºC | 60 | ||
Final extension | 72 ºC | 210 |