A | B | C | D | |
Component | Volume | Volume (1.2X) | Final conc. | |
Forward Primer (10 µM) | 0.5 μl | 1.2 μl | 0.2 µM | |
Reverse Primer (10 µM) | 0.5 μl | 1.2 μl | 0.2 µM | |
PowerPol 2X PCR Master Mix | 12.5 μl | 15 μl | - | |
ddH20 | 10.25 μl | 11.1 μl | - | |
Total volume | 23.75 μl | 28.5 μl | - |
A | B | C | D | |
Step | Temp | Sec | Cycle | |
Initial denaturation | 95 ºC | 180 | ||
Denaturation | 95 ºC | 30 | 25 cycles | |
Annealing | 60-66 ºC varied (b) | 30 | ||
Extension | 72 ºC | 180 | ||
Final extension | 72 ºC | 420 | ||
Preservation | Preservation | 4 ºC | ∞ |
A | B | C | D | |
Component | Volume | Volume (1.2X) | Final conc. | |
ZEJU PCR Master Mix | 7.5 µL | 9 µL | - | |
ddH20 | 5.55 µL | 6.66 µL | - | |
Total volume | 13.05 µL | 15.66 µL | - |
A | B | C | D | |
Step | Temp | Sec | Cycle | |
Initial denaturation | 98 ºC | 30 | ||
Denaturation | 98 ºC | 15 | 15 cycles | |
Annealing | 64-68 ºC varied (a) | 15 | ||
Extension | 72 ºC | 20 (b) | ||
Final extension | 72 ºC | 210 | ||
Preservation | Preservation | 4 ºC | ∞ |