A | B | C | |
PCR Component | Test Buffer tube (in uL) | Control buffer tube (in uL) | |
PCR grade water | Variable up to 20 uL | Variable up to 20 uL | |
dNTP mix (10 mM) | 0.4 | 0.4 | |
Thermopol buffer (10x) | / | 2 | |
Test PCR buffer (10x) | 2 | / | |
Fw/Rv primers | 1/1 | 1/1 | |
DNA template | 0.5 | 0.5 | |
DNA polymerase | 1 | 1 |
A | B | C | D | |
Phase | 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 |