A | B | C | |
25 μl RXN | FINAL CONC. | ||
Q5 Hot Start High-Fidelity 2X Master Mix | 12.5 μl | 1X | |
10 μM Forward Primer | 1.25 μl | 0.5 μM | |
10 μM Reverse Primer | 1.25 μl | 0.5 μM | |
Template DNA (1–25 ng/μl) | 1 μl | 1-25 ng | |
Nuclease-free water | 9.0 μl |
A | B | C | |
STEP | TEMP | TIME | |
Initial Denaturation | 98°C | 30 seconds | |
25 Cycles | 98°C | 10 seconds | |
50–72°C* | 10–30 seconds | ||
72°C | 20–30 seconds/kb | ||
Final Extension | 72°C | 2 minutes | |
Hold | 4–10°C |
A | B | C | |
VOLUME | FINAL CONC. | ||
PCR Product | 1 μl | ||
2X KLD Reaction Buffer | 5 μl | 1X | |
10X KLD Enzyme Mix | 1 μl | 1X | |
Nuclease-free Water | 3 μl |